Topical sanitizing formulations and uses thereof

ABSTRACT

Several embodiments disclosed herein relate to formulations having antimicrobial and/or sanitizing effects, and uses of the same. In particular, the formulations and methods of using same provide, in several embodiments, an immediate and a persistent antimicrobial effect against a broad spectrum of microorganisms.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/664,542, filed Jul. 31, 2017, which is a continuation of U.S. patentapplication Ser. No. 14/416,010, filed Jul. 19, 2013, now issued as U.S.Pat. No. 9,717,669 on Aug. 1, 2017, which is the U.S. National Stageunder 35 U.S.C. § 371 of International Application No.PCT/US2013/051398, filed Jul. 19, 2013, which claims the benefit of U.S.Provisional Application No. 61/674,779, filed on Jul. 23, 2012, theentire disclosure of which is incorporated by reference herein.

BACKGROUND Field

Several embodiments of the inventions disclosed herein relate toformulations having antimicrobial effects, and uses of the same. In someembodiments, the formulations are used to kill, inhibit, and/orotherwise reduce the number, activity and/or growth of microorganisms.

Description of the Related Art

Microorganisms (or microbes) are single cell, cell cluster, ormulticellular microscopic (or macroscopic) organisms including but notlimited to, bacteria, fungi, archaea, protists, plants (e.g., greenalgae), viruses, prions, parasites, and animals (e.g. amoeba, plankton).Microorganisms may be beneficially exploited in a variety of fields,such as biotechnology, food and beverage preparation, and diagnostictechnologies. In other contexts, however, short or long-term exposure tomicroorganisms may be harmful and lead to one or more of a variety ofillnesses and diseases in plants, animals, and humans.

SUMMARY

Contamination of surfaces with one or more types of microorganisms,and/or the transfer of microorganisms between surfaces can lead to avariety of negative effects, including but not limited to illness,disease, passage of disease, increased medical care costs, and/or lossof work time and associated loss of revenue. To address such issues andreduce the presence of a broad spectrum of microorganisms from a varietyof surfaces, there is provided herein a method of dual-action immediateand persistent microbial reduction, the method comprising, applying atopical formulation to a surface contaminated with both gram-positiveand gram-negative bacteria. In one embodiment, the topical formulationcomprises, consists essentially of, or consists of a terpene, tocopherylacetate, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride, wherein the formulation kills (or otherwiseinactivates) at least 90% (e.g., 99%) of both gram-positive andgram-negative bacteria within 45 seconds (e.g., 20 seconds) of contactwith the microbes. The formulation is configured to kill (or otherwiseinactivate) at least 85% (e.g., 90%) of both gram-positive andgram-negative bacteria at least 4 hours post-application of theformulation in one embodiment. In one embodiment, the effect persistsfor at least 6 or 8 hours post-application. Persistence, according tosome embodiments, is achieved by a non-evaporative component of theformulation. Advantageously, in several embodiments, an antimicrobialcomponent remains on the surface (hands, equipment, etc.) that detersagainst any subsequent contamination. This persistent antimicrobialbarrier may be physical, chemical or both, and persists, in someembodiments, even after the surface is rinsed. Thus, in one embodiment,the persistent barrier is water-proof or water-resistant.

Triclosan-free formulations are provided in several embodiments. Manyembodiments disclosed herein are particularly advantageous because theyare efficacious without contributing to microbial drug resistance.

In several embodiments, a method of dual-action immediate and persistentmicrobial reduction is provided. In one embodiment, the method comprisesapplying a topical formulation to a surface contaminated with aplurality of microbes. In one embodiment, the topical formulationcomprises, consists essentially of or consists of a terpene, tocopherylacetate, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride, wherein the formulation kills (or otherwiseinactivates) at least 90% (e.g., 99%) of the plurality of microbeswithin 45 seconds (e.g., 20 seconds) of contact with the microbes. Inone embodiment, the formulation is configured to kill (or otherwiseinactivate) at least 85% (e.g., 90%) of the plurality of microbes atleast 4 hours post-application of the formulation, wherein the pluralityof microbes comprises two or more of the following: gram-positivebacteria, gram-negative bacteria, fungi, mold, yeast and viruses. In oneembodiment, the effect persists for at least 6 or 8 hourspost-application.

The methods disclosed herein are useful for providing antimicrobialeffects to a variety of surfaces. In several embodiments, the surface ishuman or animal skin or an inorganic surface (including but not limitedto countertops, door handles, faucets, telephones beds/bed frames, bedlinens, medical equipment, computers, writing instruments, surgicalequipment). In several embodiments, the surface is human or animal skin,and wherein the application does not induce skin irritation.Advantageously, in several embodiments, the formulations not onlyprovide antimicrobial effects, but also moisturize, soften, and/orimprove the health of the human and/ or animal skin to which they areapplied. In several embodiments, the formulations are also used tofacilitate wound healing (e.g., by reducing or preventing infection). Inseveral embodiments, the formulations are used to treat acne, which inseveral embodiments, is particularly advantageous because the certainembodiments of the formulations disclosed herein moisturize the skin(e.g., are without drying effects). In several embodiments, fungalinfections can be treated and/or prevented by application of theformulations disclosed herein.

In several embodiments, the topical formulation dries within about 5 toabout 30 seconds. In some embodiments, shorter or longer drying timesare provided (e.g., about 5 to about 10 seconds, about 10 to about 15seconds, about 15 to about 20 seconds, about 20 to about 25 seconds,about 25 to about 30 seconds, and overlapping ranges thereof). In someembodiments, the drying time assists in the “feel” of the formulation onthe skin, in that, for example, there is a reduction in the residue orsticky, greasy, or otherwise unclean feeling that other formulations mayleave on a surface.

In several embodiments, the bacteria comprise transient bacteria, whilein other embodiments the formulations can be adjusted to provideantimicrobial effects against certain resident bacteria.

In several embodiments, the terpene is present in the topicalformulation in an amount ranging from about 0.25% to about 0.60% byweight of the formulation. In some embodiments, the tocopheryl acetateis present in the topical formulation in an amount ranging from about0.05% to about 0.5% by weight of the formulation. In some embodiments,the polyaminopropyl biguanide is present in an amount ranging from about0.05% to about 0.20% by weight of the formulation. In some embodiments,the at least one quaternary ammonium salt is present in an amountranging from about 0.01% to about 0.40% by weight of the formulation. Inseveral embodiments, the quaternary ammonium salt comprises benzethoniumchloride. Benzalkonium chloride, or another quaternary ammonium salt,may be used instead of or in addition to benzethonium chloride in someembodiments.

In several embodiments, the formulation further comprises alcohol in anamount ranging from about 8% to about 20% by weight of the formulation.In several embodiments the formulation may also further comprisechlorhexidine gluconate in an amount ranging from about 0.10% to about0.40% by weight of the formulation. The formulation, depending on theembodiment may also further comprise hydroxyethyl ethylcellulose in anamount ranging from about 0.05% to about 0.20% by weight of theformulation. In some embodiments, the formulation further comprises oneor more zinc salts in an amount ranging from about 0.05% to about 0.60%by weight of the formulation. In some embodiments, the zinc saltcomprises zinc lactate, zinc gluconate, combinations thereof, and/or azinc matrix. In several embodiments, the formulation further comprisespanthenol in an amount ranging from about 0.25% to about 0.60% by weightof the formulation. In several embodiments, the terpene comprises asesquiterpene, and in certain such embodiments, the sesquiterpenecomprises farnesol. Other terpenes, or combinations of terpenes (and/ortheir derivatives) are used in some embodiments.

In several embodiments, the formulation further comprises one or moreadditional compounds, such as emollients. In some embodiments, theformulation further comprises one or more of Vitamins A, B, K, C, andaloe. In several embodiments, the topical formulation further comprisesan essential oil, such as for example, eucalyptus oil. In someembodiments, the eucalyptus oil is present in an amount ranging fromabout 0.05% to about 0.5% by weight of the formulation. Other essentialoils, alone or in combination, may also be used.

In several embodiments, application of the topical formulation comprisesdispensing between about 0.1 and about 0.8 mL (e.g., about 0.1 mL andabout 0.2 mL, about 0.2 mL and about 0.3 mL, about 0.3 mL and about 0.4mL, about 0.4 mL and about 0.5 mL, about 0.5 mL and about 0.6 mL, about0.6 mL and about 0.7 mL, about 0.7 mL and about 0.8 mL, and overlappingranges thereof) of the formulation onto the surface. Advantageously, inseveral embodiments, the efficacy of the formulation at 4 hours (ormore) post-application of the formulation is achieved in the absence ofreapplication of the formulation to the surface.

There is further provided, in several embodiments, a method ofdual-action immediate and persistent microbial reduction, the methodcomprising applying a topical formulation to a surface contaminated witha plurality of microbes, the topical formulation comprising a terpene,polyaminopropyl biguanide and at least one quaternary ammonium saltselected from the group consisting of benzethonium chloride andbenzalkonium chloride (or a combination thereof), wherein theformulation kills at least 99% of the plurality of microbes within 20seconds (e.g., within 2, 5, 10, 15 seconds) of contact with themicrobes, wherein the formulation is configured to kill at least 90% ofthe plurality of microbes at least 4 hours (or more) post-application ofthe formulation, and wherein the plurality of microbes comprises two ormore of the following: gram-positive bacteria, gram-negative bacteria,fungi, and viruses.

Moreover, there is provided a method of dual-action immediate andpersistent microbial reduction, the method comprising, applying atopical formulation to a surface contaminated with both gram-positiveand gram-negative bacteria, wherein the topical formulation comprises aterpene, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride, wherein the formulation kills at least 99% of theplurality of microbes within 20 seconds (e.g., within 2, 5, 10, 15seconds) of contact with the microbes, wherein the formulation isconfigured to kill at least 95% of the plurality of microbes at least 4hours (or more) post-application of the formulation, and wherein theformulation is configured to kill at least 90% of both gram-positive andgram-negative bacteria at least 4 hours (or more) post-application ofthe formulation. There is additionally provided a method of dual-actionimmediate and persistent microbial reduction, the method comprising,applying a topical formulation to a surface contaminated with aplurality of microbes, wherein the topical formulation comprises aterpene, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride, wherein the formulation kills at least 99% of theplurality of microbes within 20 seconds of contact with the microbes,wherein the formulation is configured to kill at least 90% of theplurality of microbes at least 4 hours post-application of theformulation, and wherein the plurality of microbes comprises two or moreof the following: gram-positive bacteria, gram-negative bacteria, fungi,and viruses.

In several embodiments, the topical formulation further comprisesalcohol in an amount ranging from about 60% to about 90% by weight ofthe formulation. In several embodiments, the formulation is suitable forapplication to a surface that is human or animal skin or an inorganicsurface. In those cases where the is human or animal skin, and whereinthe application does not induce skin irritation, which includes but isnot limited to drying, cracking, reddening, itchiness, burning or otheradverse sensations.

In several embodiments, the terpene is present in the topicalformulation in an amount ranging from about 0.25% to about 0.60% byweight of the formulation. In some embodiments, the tocopheryl acetateis present in the topical formulation in an amount ranging from about0.05% to about 0.5% by weight of the formulation. In some embodiments,the polyaminopropyl biguanide is present in an amount ranging from about0.05% to about 0.20% by weight of the formulation. In some embodiments,at least one quaternary ammonium salt is present in an amount rangingfrom about 0.01% to about 0.40% by weight of the formulation. In severalembodiments, the quaternary ammonium salt comprises benzethoniumchloride. Benzalkonium chloride and/or another quaternary ammonium saltmay be used in additional embodiments.

In several embodiments, the formulation further comprises alcohol in anamount ranging from about 8% to about 20% by weight of the formulation.In several embodiments the formulation may also further comprisechlorhexidine gluconate in an amount ranging from about 0.10% to about0.40% by weight of the formulation. The formulation, depending on theembodiment may also further comprise hydroxyethyl ethylcellulose in anamount ranging from about 0.05% to about 0.20% by weight of theformulation. In some embodiments, the formulation further comprises oneor more zinc salts in an amount ranging from about 0.05% to about 0.60%by weight of the formulation. In some embodiments, the zinc saltcomprises zinc lactate, zinc gluconate, combinations thereof, and/or azinc matrix. In several embodiments, the formulation further comprisespanthenol in an amount ranging from about 0.25% to about 0.60% by weightof the formulation. In several embodiments, the terpene comprises asesquiterpene, and in certain such embodiments, the sesquiterpenecomprises farnesol. Other terpenes, or combinations of terpenes (and/ortheir derivatives) are used in some embodiments.

In several embodiments, the formulation further comprises one or moreadditional emollient compounds. In some embodiments, the formulationfurther comprises one or more of Vitamins A, B, K, E, and C. In severalembodiments, the topical formulation further comprises an essential oil,such as for example, eucalyptus oil. In some embodiments, the eucalyptusoil is present in an amount ranging from about 0.05% to about 0.5% byweight of the formulation. Other essential oils, alone or incombination, may also be used.

In several embodiments, application of the topical formulation comprisesdispensing between about 0.1 mL and about 0.8 mL (e.g., about 0.1 mL andabout 0.2 mL, about 0.2 mL and about 0.3 mL, about 0.3 mL and about 0.4mL, about 0.4 mL and about 0.5 mL, about 0.5 mL and about 0.6 mL, about0.6 mL and about 0.7 mL, about 0.7 mL and about 0.8 mL, and overlappingranges thereof) of the formulation onto the surface. Advantageously, inseveral embodiments, the efficacy of the formulation at 2-8 hours (e.g.,6 hours) post-application of the formulation is achieved in the absenceof reapplication of the formulation to the surface. In severalembodiments, efficacy of the formulation persists for longer periods oftime, even in the absence of reapplication.

There is additionally provided a method of immediate microbialreduction, the method comprising applying a topical formulation to asurface contaminated with both gram-positive and gram-negative bacteria,wherein the topical formulation comprises a terpene, tocopheryl acetate,polyaminopropyl biguanide and at least one quaternary ammonium saltselected from the group consisting of benzethonium chloride andbenzalkonium chloride, wherein the formulation kills at least 90% ofboth gram-positive and gram-negative bacteria within 20 seconds (e.g.,within 2, 5, 10, 15 seconds) of contact with the microbes. Further,there is provided a method of immediate microbial reduction, the methodcomprising, applying a topical formulation to a surface contaminatedwith a plurality of microbes, wherein the topical formulation comprisesa terpene, tocopheryl acetate, polyaminopropyl biguanide and at leastone quaternary ammonium salt selected from the group consisting ofbenzethonium chloride and benzalkonium chloride, wherein the formulationkills at least 90% of the plurality of microbes within 20 seconds (e.g.,within 2, 5, 10, 15 seconds) of contact with the microbes, and whereinthe plurality of microbes comprises two or more of the following:gram-positive bacteria, gram-negative bacteria, fungi, and viruses.

In still additional embodiments, there are provided method for theimmediate and persistent reduction in microbial populations, the methodscomprising applying an alcohol-based topical formulation to a surfacecontaminated with both gram-positive and gram-negative bacteria, whereinthe topical formulation comprises at least about 65% ethyl alcohol byweight, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride. In several embodiments, the application of theformulation results in a reduction of least 95% (e.g., 99%) of bothgram-positive and gram-negative bacteria within 45 seconds (e.g., 20seconds) of contact with the bacteria, and wherein application of theformulation also result in a reduction of at least 80% (e.g., 90%, 95%)of both gram-positive and gram-negative bacteria at least 2 hours (e.g.,3 hours, 4 hours) post-application of the formulation. In severalembodiments, the significant efficacy persists between the immediate andpersistent time frames, e.g., there is significant antimicrobialefficacy at about 10 minutes, about 20 minutes, about 25 minutes, about30 minutes, about 35 minutes, about 40 minutes, about 60 minutes, orabout 90 minutes (and times therebetween) after application.

In several embodiments, there are also provided methods for preventingbacterial colonization of a surface (such as human skin), the methodcomprising applying a topical formulation to a surface susceptible tocontamination with both bacteria, wherein the topical formulationcomprises a terpene, tocopheryl acetate, polyaminopropyl biguanide,chlorhexidine digluconate, and at least one quaternary ammonium saltselected from the group consisting of benzethonium chloride andbenzalkonium chloride. In several embodiments, the formulation kills atleast 99% of the bacteria within 20 seconds of contact with the microbes(resulting in short-term prevention of colonization) and kills at least85% of both gram-positive and gram-negative bacteria at least 8 hourspost-application of the formulation (resulting in prevention ofcolonization on a long-term time frame). In some embodiments, theformulation kills at least 95% of the bacteria within about 45 secondsof contact with the microbes (resulting in short-term prevention ofcolonization) and kills at least 80% of both gram-positive andgram-negative bacteria at least 6, 8, 12, or 24 hours post-applicationof the formulation (resulting in prevention of colonization on along-term time frame). In other words, gram-positive and gram-negativebacteria continue to be inhibited even after 6 hours post application ofthe formulation. In some embodiments, this time period is lengthened to,for example, 8 hours, 12 hours or 24 hours.

In several embodiments, the persistence of the formulations is due to,at least in part, the continued presence of the formulations on asurface (such as the hands of a user). In several embodiments, thecontinued presence of the formulation is due to chemical or physicalinteractions between the components of the formulation and the surface.For example, in several embodiments wherein the formulations are appliedto the skin of a user, the formulation ionically binds to the skin ofthe user. Thus, the formulation is present on the skin of the user for alonger period of time than, for example many alcohol based productswhose efficacy is significantly, if not completely, diminished uponevaporation of the alcohol. In several embodiments, reversible covalentbonding occurs between the formulation and the skin of the user.Advantageously, in several embodiments, the continued presence of theformulations is not associated with adverse feel or residue (e.g., nosticky or greasy hand feel). Moreover, in several embodiments, theformulations are configured to remain on the surface after exposure ofthe surface to conditions that would result in the substantial removalof other products. For example, in several embodiments, even aftermultiple (e.g., 3, 4, 5, 6, or more) hand washings, a significantpercentage of the formulations remain on the hands of a user (e.g., atleast about 80%, about at least 70%, at least about 60 percent, at leastabout 50%, and percentages therebetween). Advantageously, in severalembodiments, even if the handwashing does result in reduction in thepresence of the formulations on a surface (e.g., the hands of the user),the efficacy of the formulation persists. For example, in severalembodiments, reduction in the amount of the formulation that remains ona subject's hands can be reduced by about 20%, about 40%, about 60%,about 80% (or greater) while substantial efficacy of the formulation ismaintained. Thus, in several embodiments, the persistence allows theformulations to remain present on the surface that has been treated,such that the surface is resistant to microbial colonization for anextended period of time, even without reapplication of the formulations.In several embodiments, the formulations can be reapplied, which resultsin further protection of the surface.

In several embodiments, there are additionally provided methods oftreating a surface colonized with microbes by applying a topicalformulation to a colonized surface, wherein the topical formulationcomprises a terpene, polyaminopropyl biguanide, a zinc salt, and atleast one quaternary ammonium salt selected from the group consisting ofbenzethonium chloride and benzalkonium chloride. Treatment of thesurface with the formulation, in several embodiments, results in atleast a 95% (e.g., 99%) reduction of the microbe population within 45seconds (e.g., 20 seconds) of contact with the microbes, andsurprisingly, continues to result in at least 90% reduction in themicrobe population at time points of at least 4 hours post-applicationof the formulation. In some embodiments, the reduction of 90% reductionis also seen at 8 hours, 12 hours or 24 hours.

In several embodiments of the formulations described herein, microbialreduction levels drop by 1-5% every hour. As an example, if a 99%reduction in microbes is observed at hour 0 (immediately postapplication of the formulation), a 98% reduction is seen at hour 1, a97% reduction is seen at hour 2, a 96% reduction is seen at hour 3, a95% reduction is seen at hour 4, etc. In one embodiment, this reductiontrend lasts for the first 12, 18 or 24 hours. The percent reduction iscompared to the number and/or function of microbes pre-application of anantimicrobial formulation described herein.

In view of the continued development of resistance to antibiotics ofcertain microbe populations, there is also advantageously providedherein methods for the dual-action immediate and persistent reduction ofantibiotic resistant bacteria, the method comprising applying a topicalformulation to a surface contaminated with antibiotic resistantbacteria, wherein the topical formulation comprises a terpene,polyaminopropyl biguanide, a zinc salt, and at least one quaternaryammonium salt selected from the group consisting of benzethoniumchloride and benzalkonium chloride. Application of the formulationresults in a short term (e.g., within about 120 seconds) kill of atleast 99% of the antibiotic resistant bacteria, and a long-term (e.g.,for at least about 6 hours after application of the formulation) kill ofat least 80% of the antibiotic resistant bacteria.

Also provided is a method of reduction of antibiotic resistant bacteriaon a short and long-term basis comprising applying a topical formulationto a surface contaminated with antibiotic resistant bacteria, whereinthe topical formulation comprises at least about 65% ethyl alcohol byweight, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride. Application of the formulation results in a shortterm (e.g., within about 120 seconds) kill of at least 99% of theantibiotic resistant bacteria, and a long-term (e.g., for at least about6 hours after application of the formulation) kill of at least 80% ofthe antibiotic resistant bacteria. Additionally provided are methods ofreduction of drug-resistant viruses on a short and long-term basiscomprising applying a formulation to a surface contaminated withdrug-resistant viruses, wherein the topical formulation comprises ethylalcohol, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride. In still additional embodiments, the topicalformulations disclosed herein are used to eliminate viruses from asurface, without inducing resistance to the formulation in the viruses.

Moreover, several embodiments relate to methods of reducing bacterialcontamination of a surface, without intruding bacterial resistance, themethod comprising applying a topical formulation to a surface each timethat surface is contaminated with the bacteria, wherein the topicalformulation comprises a terpene, polyaminopropyl biguanide and at leastone quaternary ammonium salt selected from the group consisting ofbenzethonium chloride and benzalkonium chloride, wherein the formulationkills at least 90% of the bacteria after a first contamination, andwherein the formulation kills at least 99.9% of the bacteria after atleast 9 additional contaminations of the surface. Advantageously suchformulations and methods can be used to repeatedly decontaminate asurface of microbes, without loss of efficacy against the microbes,despite repeated applications. Additionally, several embodiments relateto methods of reducing viral contamination of a surface, withoutintruding viral drug resistance, the method comprising applying atopical formulation to a surface each time that surface is contaminatedwith the virus (or viruses), wherein the formulation comprises aterpene, polyaminopropyl biguanide and at least one quaternary ammoniumsalt selected from the group consisting of benzethonium chloride andbenzalkonium chloride. As above, advantageously such formulations andmethods can be used to repeatedly decontaminate a surface of virus (evenof distinct viral types), without loss of efficacy against the viruses,despite repeated applications. Such methods are of particular importancein environments where repeated contamination is possible, such as, forexample, health care facilities. Additionally, the methods andformulations (discussed in more detail below) can be used in otherarenas, such as for example, schools or workplaces (to reduceabsenteeism), food preparation areas (to reduce food contamination), aswell as other environments where microbial contamination is possible.Moreover, in several embodiments, the efficacy of the formulations issuch that smaller amounts may be used to generate efficacy equivalent tothat of other products (which require larger application amounts). Thus,several embodiments of the formulations provide effective protectionagainst microbial contamination, but at significant cost savings.

In several embodiments, the formulations described herein are configuredin a form selected from the group consisting of a powder, spray,sanitizer, scrub, gel, lotion, cream, foaming soap, and block soap. Inadditional embodiments, the formulation is suitable for incorporationinto an additional formulation selected from the group consisting ofbody wash, shampoo, conditioner, and cosmetic base. Although severaltopical formulations are provided, non-topical formulations are alsoprovided (e.g., oral, systemic, inhalants, etc.). In some embodiments,formulations described herein are embedded within or coated on tomaterials. Antimicrobial wipes are provided in several embodiments. Insome embodiments, wipes comprise a single layer or multiple layers thatare woven or non-woven. Materials include but are not limited to naturalor synthetic materials such as cotton, soya, hemp, rayon, polyester,nylon or other polymers. The wipes can be wet or dry. Wipes (or sponges)may be provided with or without a handle. Wipes may include several ofthe formulations described herein. Wipes may be suitable for skin orsurfaces (floors, counters, lavatories). For human use, wipes mayinclude emollient or other dermatologically beneficial ingredients.

In addition to the methods and processes disclosed herein, there arealso provided antimicrobial formulations that provide unexpectedlyadvantageous antimicrobial effects, on an immediate time frame, apersistent time frame, and/or an immediate and a persistent time frame.Thus, there is provided, in several embodiments, a dual-actionantimicrobial formulation for both immediate and persistent reduction ofgram-positive and gram-negative bacteria, the formulation comprising,consisting essentially of or consisting of a quaternary ammonium salt inan amount ranging from about 0.10% to about 0.40% by weight of theformulation, wherein the quaternary ammonium salt comprises benzethoniumchloride, benzalkonium chloride, or combinations thereof, a terpene orderivative thereof in an amount ranging from about 0.25% to about 0.60%by weight of the formulation, tocopheryl acetate in an amount rangingfrom about 0.05% to about 0.5% by weight of the formulation,polyaminopropyl biguanide in an amount ranging from about 0.05% to about0.20% by weight of the formulation, wherein the formulation kills atleast 99% of both gram-positive and gram-negative bacteria within 20seconds of contact with the microbes, and wherein the formulation isconfigured to kill at least 90% of both gram-positive and gram-negativebacteria at least 4 hours post-application of the formulation. Inseveral embodiments, the formulations comprise, consist of, or consistessentially of natural ingredients (e.g., non-synthetic). Combinationsof natural and synthetic ingredients are used in additional embodiments.

There is also provided a dual-action antimicrobial formulation for bothimmediate and persistent reduction of broad spectrum microbes, theformulation comprising, consisting essentially of or consisting of aquaternary ammonium salt in an amount ranging from about 0.10% to about0.40% by weight of the formulation, wherein the quaternary ammonium saltcomprises benzethonium chloride, benzalkonium chloride, or combinationsthereof, a terpene or derivative thereof in an amount ranging from about0.25% to about 0.60% by weight of the formulation, tocopheryl acetate inan amount ranging from about 0.05% to about 0.5% by weight of theformulation, polyaminopropyl biguanide in an amount ranging from about0.05% to about 0.20% by weight of the formulation, wherein theformulation kills at least 99% of a plurality of microbes within 20seconds of contact with the microbes, wherein the formulation isconfigured to kill at least 90% of the plurality of microbes at least 4hours post-application of the formulation, and wherein the plurality ofmicrobes comprises two or more of the following: gram-positive bacteria,gram-negative bacteria, fungi, and viruses.

In several embodiments, a sanitizing gel or non-gel formulation isprovided comprising, consisting essentially of, or consisting of (byweight of the formulation) deionized water in an amount ranging fromabout 20% to about 30% (e.g., 26.98%), D,L panthenol in an amountranging from about 0.2% to about 2.0% (e.g., 1.0%), Structure CEL(comprising hydroxyethyl ethylcellulose, water, sodium sulfate, sodiumcitrate, BHT) in an amount ranging from about 0.4% to about 1.0% (e.g.,0.7%), 200 proof denatured alcohol in an amount ranging from about 60%to about 75% (e.g., 69.0%), PEG-12 dimethicone in an amount ranging fromabout 0.1% to about 0.3% (e.g., 0.2%), farnesol in an amount rangingfrom about 0.1% to about 1.0% (e.g., 0.5%), benzethonium chloride in anamount ranging from about 0.05% to about 0.2% (e.g., 0.12%), andcosmocil CQ (comprising water and polyaminopropyl biguanide) in anamount ranging from about 0.5% to about 3% (e.g., 1.5%).

In several embodiments, a non-gel sanitizer lotion is providedcomprising, consisting essentially of or consisting of (by weight of theformulation) deionized water in an amount ranging from about 60% toabout 75% (e.g., 69.36%), polyquaternium-10 in an amount ranging fromabout 0.05% to about 1.0% (e.g., 0.1%), glycerine in an amount rangingfrom about 1.5% to about 3.5% (e.g., 2.75%), D,L panthenol in an amountranging from about 0.1% to about 2% (e.g., 0.5%), zinc gluconate in anamount ranging from about 0.1% to about 2% (e.g., 0.3%), incroquatbehenyl TMS in an amount ranging from about 3.0% to about 5.0% (e.g.,4.2%), stearamidopropyl dimethylamine in an amount ranging from about0.5% to about 2% (e.g., 0.84%), cetearyl alcohol in an amount rangingfrom about 3.0% to about 5.0% (e.g., 3.8%), dimethicone in an amountranging from about 0.5% to about 2.0% (e.g., 1.0%), farnesol in anamount ranging from about 0.1% to about 2% (e.g., 0.5%), tocopherylacetate in an amount ranging from about 0.01 to about 1.0% (e.g., 0.1%);citric acid in an amount ranging from about 0.01% to about 1.0% (e.g.,0.15%), benzethonium chloride in an amount ranging from about 0.1% toabout 1.5% (e.g., 0.2%); euxyl K 700 (comprising benzyl alcohol,phenoxyethanol, potassium sorbate, tocopherol) in an amount ranging fromabout 0.1% to about 1.5% (e.g., 0.5%), 200-proof alcohol in an amountranging from about 10.0% to about 20.0% (e.g., 14.0%), chlorhexidinedigluconate in an amount ranging from about 0.1% to about 1.0% (e.g.,0.2%), and cosmocil CQ (comprising water and polyaminopropyl biguanide)in an amount ranging from about 0.05% to about 3.0% (e.g., 1.5%).

In several embodiments, a non-gel sanitizer is provided. Many of theformulations described herein are provided in non-gel formats, and areparticularly advantageous in being more liquid or more solid thantypical gels.

In several embodiments, a surgical scrub is provided comprising (byweight of the formulation) deionized water in an amount ranging fromabout 20.0% to about 30.0% (e.g., 27.68%), D,L-panthenol in an amountranging from about 0.5% to about 2.0% (e.g., 1.0%), PEG-12 dimethiconein an amount ranging from about 0.05% to about 0.5% (e.g., 0.2%),farnesol in an amount ranging from about 0.1% to about 2.0% (e.g.,0.5%), 200 proof alcohol in an amount ranging from about 60% to about75% (e.g., 69.0%), cosmocil CQ (comprising water and polyaminopropylbiguanide) in an amount ranging from about 0.5% to about 3.0% (e.g.,1.5%), and benzethonium chloride in an amount ranging from about 0.05%to about 1.0% (e.g., 0.12%).

In several embodiments, a persistent foaming soap is provided comprising(by weight of the formulation) deionized water in an amount ranging fromabout 50.0% to about 65.0% (e.g., 64.02%), polyquaternium-10 in anamount ranging from about 0.005% to about 0.1% (e.g., 0.01%), citricacid in an amount ranging from about 0.1% to about 2.0% (e.g., 0.7%),butylene glycol in an amount ranging from about 0.5% to about 2.0%(e.g., 1.0%), zinc gluconate in an amount ranging from about 0.1% toabout 2.0% (e.g., 0.3%), benzethonium chloride in an amount ranging fromabout 0.05% to about 2.0% (e.g., 0.2%), mackamine LO (comprising waterand lauramine oxide) in an amount ranging from about 5.0% to about 10.0%(e.g., 8.0%), caltaine C-35 (comprising water cocamiopropyl betaine, andsodium chloride) in an amount ranging from about 5.0% to about 10.0%(e.g., 7.0%), carsoquat CT-429 (comprising water and cetrimoniumchloride) in an amount ranging from about 3.0% to about 10.0% (e.g.,5.0%), PPG-2 hydroxyethyl cocamide in an amount ranging from about 1.0%to about 5.0% (e.g., 3.0%), methylchloroisothiazolinone in an amountranging from about 0.01% to about 1.0% (e.g., 0.07%), actiphyte of aloevera (comprising glycerin, water aloe barbadensis leaf extract) in anamount ranging from about 0.05% to about 1.0% (e.g., 0.1%), 200 proofalcohol in an amount ranging from about 5.0% to about 20.0% (e.g.,10.0%), farnesol in an amount ranging from about 0.1% to about 1.0%(e.g., 0.5%), fragrance (for example citrus fragrance) in an amountranging from about 0.01 to about 0.10% (e.g., 0.05%), and cosmocil CQ(comprising water and polyaminopropyl biguanide) in an amount rangingfrom about 0.01% to about 0.5% (e.g., 0.05%).

In several embodiments, a non-persistent foaming soap is providedcomprising (by weight of the formulation) deionized water in an amountranging from about 60% to about 70% (e.g., 68.58%), butylene glycol inan amount ranging from about 0.5% to about 2.0% (e.g., 1.0%), citricacid in an amount ranging from about 0.1% to about 1.0% (e.g., 0.6%),zinc gluconate in an amount ranging from about 0.1% to about 1.0% (e.g.,0.3%), benzethonium chloride in an amount ranging from about 0.05% toabout 1.0% (e.g., 0.2%), mackamine LO (comprising water and lauramineoxide) in an amount ranging from about 5.0% to about 10.0% (e.g., 8.0%),caltaine C-35 (comprising water cocamiopropyl betaine, and sodiumchloride) in an amount ranging from about 5.0% to about 10.0% (e.g.,7.0%), carsoquat CT-429 (comprising water and cetrimonium chloride) inan amount ranging from about 3.0% to about 10.0% (e.g., 5.0%), lauramideDEA in an amount ranging from about 0.5% to about 2.0% (e.g., 1.1%),PPG-2 hydroxyethyl cocamide in an amount ranging from about 1.0% toabout 5.0% (e.g., 3.0%), methylchloroisothiazolinone in an amountranging from about 0.01% to about 1.0% (e.g., 0.07%), actiphyte of aloevera (comprising glycerin, water aloe barbadensis leaf extract) in anamount ranging from about 0.05% to about 1.0% (e.g., 0.1%), 200 proofalcohol in an amount ranging from about 2.0% to about 20.0% (e.g.,5.0%), and fragrance (for example natural fragrances, such as a menthol,lavender or citrus fragrance) in an amount ranging from about 0.01 toabout 0.10% (e.g., 0.05%). In one embodiment, the fragrance contributesto the antimicrobial properties and/or other desirable effect

In several embodiments, the formulation is suitable for application to asurface is infected gram-positive and gram-negative bacteria and/or aplurality of microbes, such as for example human or animal skin or aninorganic surface.

In several embodiments, the terpene is present in the topicalformulation in an amount ranging from about 0.25% to about 0.60% byweight of the formulation. In several embodiments, the terpene orderivative thereof comprises farnesol. In some embodiments, anotherterpenoid or derivative thereof is used in addition to, or in place offarnesol. In several embodiments, tocopheryl acetate is present in thetopical formulation in an amount ranging from about 0.05% to about 0.5%by weight of the formulation. In several embodiments, polyaminopropylbiguanide is present in an amount ranging from about 0.05% to about0.20% by weight of the formulation. In several embodiments, thequaternary ammonium salt comprises benzethonium chloride.

In several embodiments, the formulation further comprises alcohol in anamount ranging from about 8% to about 20% by weight of the formulation.Additionally, in several embodiments, the topical formulation furthercomprises chlorhexidine gluconate in an amount ranging from about 0.10%to about 0.40% by weight of the formulation. In some embodiments, theformulation further comprises hydroxyethyl ethylcellulose in an amountranging from about 0.05% to about 0.20% by weight of the formulation. Inseveral embodiments, the formulation further comprises one or more zincsalts in an amount ranging from about 0.05% to about 0.60% by weight ofthe formulation. In some embodiments, the one or more zinc saltscomprises zinc lactate, zinc gluconate, combinations thereof, and/or azinc matrix. In several embodiments, the formulation further comprisespanthenol in an amount ranging from about 0.25% to about 0.60% by weightof the formulation.

In several embodiments, the formulation further comprises one or moreadditional emollient compounds. In several embodiments, the formulationfurther comprises one or more of Vitamins A, B, K, and C. In someembodiments, the formulation further comprises an essential oil. In someembodiments, the essential oil comprises eucalyptus oil. In severalembodiments, the eucalyptus oil is present in an amount ranging fromabout 0.05% to about 0.5% by weight of the formulation. Other oils maybe used, depending on the embodiment. In some embodiments, combinationsof oils are used, either alone or in combination with a fragrance, inorder to provide a desirable scent or odor to the formulation.

In several embodiments, the dual action antimicrobial effects areachieved by dispensing between about 0.1 and 0.8 mL of the formulationonto a surface infected with gram-positive and gram-negative bacteriaand/or a plurality of microbes. In several embodiments, the amountrequired to achieve immediate and/or persistent effects is on the lowerend of the range. This is advantageous in several embodiments, becausethis increases the usability of the formulation (e.g., a wider range ofusers can correctly use the product, such as small children, individualswith limited flexibility, etc.). In some embodiments, the larger volumesare advantageous, however, as they allow complete coverage of certainsurfaces (e.g., a surgical surface or equipment). In conjunction withthe viscosities of the formulations disclosed herein, achieving highlyeffective, broad spectrum antimicrobial effects is more easily achievedin several embodiments. While multiple doses are desired in someembodiments, in several embodiments, the immediate and/or the persistentantimicrobial effects are achieved using a single dose of theformulation. In some embodiments, healthy microbes (e.g., good bacteria)are unaffected or less affected by some of the formulations describedherein.

In several embodiments, there is also provided a kit comprising one ormore of the formulations disclosed herein as well as instructions foruse of the particular formulation. In some embodiments, kits maycomprise applicators.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts data comparing the antimicrobial effectiveness of varioussanitizing formulations after multiple re-infections with S. marcescensand multiple applications of the formulations.

FIG. 2 depicts summary data related to antimicrobial effectiveness ofvarious sanitizing formulations after multiple re-infections with S.marcescens and multiple applications of the formulations.

FIG. 3 depicts data related to the antimicrobial effectiveness ofvarious sanitizing formulations disclosed herein against S. marcescensas compared to FDA recommended levels of efficacy after multipleapplications.

FIG. 4 depicts data comparing the antimicrobial effectiveness (againstS. marcescens) of one embodiment of an alcohol-based sanitizingformulation disclosed herein as compared to one embodiment of anantiseptic lotion formulation disclosed herein.

FIG. 5 depicts data related to the sustained antimicrobial effectivenessof one embodiment of an alcohol-based antiseptic against either E. colior S. aureus.

FIG. 6 depicts data related to the sustained antimicrobial effectivenessof one embodiment of an antiseptic lotion against either E. coli or S.aureus.

FIG. 7 depicts data related to the sustained antimicrobial effectivenessof various soaps, including one embodiment of a foaming soap disclosedherein, against E. coli.

FIG. 8 depicts data related to the sustained antimicrobial effectivenessof various soaps, including one embodiment of a foaming soap disclosedherein, against MRSA.

FIG. 9 depicts data related to the sustained antimicrobial effectivenessof one embodiment of an alcohol-based sanitizing formulation disclosedherein as compared to one embodiment of an antiseptic lotion formulationdisclosed herein, against MRSA.

DETAILED DESCRIPTION

Hospitals, ambulances, emergency treatment settings, workplaceenvironments, public places and even household settings are linked bythe ever-present threat of microorganisms, and the possibility ofdeveloping and propagating an infection, disease or other illness. Ascan be appreciated by simply viewing historical data from the Centersfor Disease Control (CDC) depicting the spread of the flu virus overseveral past years, the rate and breadth to which an illness can spreadis surprising. This is largely due to, at least in some part, the easewith which certain microorganisms can be spread. By way of example,those infected with a respiratory virus, for example, (who may not yetshow overt symptoms of the infection) can spread the virus up to adistance of about 6 feet. With respect to the flu virus (and othermicroorganisms) spreading can be due to virus-containing droplets formedwhen infected individuals cough, sneeze or talk. These droplets canunknowingly be inhaled by people who are nearby or ingested by touchinga surface or object that has flu virus on it and then touching their ownmouth or nose. Given that the number of people or surfaces an infectedindividual could come into contact with on a daily basis (for exampleriding a public bus on the way to work or out to lunch), the potentialfor spreading of microorganism-borne illnesses is vast.

For example, according to the CDC, from 1976 to 2006, estimates ofinfluenza-associated deaths in the United States alone range from a lowof about 3,000 to a high of about 49,000 people. On average, more than200,000 people are hospitalized in the United States each year due tocomplications related to seasonal influenza. The immune systems ofhealthy individuals often successfully combat such microorganism-borneillnesses, but even healthy individuals can be at heightened risk.Elderly, those who are already sick or otherwise immune-compromised, andchildren are also at risk, and perhaps even more severe outcomes. Thus,when expanded to other microorganism-borne illnesses and considering thevarious at-risk populations (especially those in less developed regionsthat lack ready access to clean water and soaps), the potential forincreased health care costs (which of course can lead to a feed-forwardspread of an illness), lost business or personal revenue, reducededucation time and productivity, and other effects, there is a need toprovide formulations and methods that can reduce and/or eliminate a widevariety of microorganisms on both a short and long-term basis. Theformulations and methods presented herein address this need by providingwide-spectrum antimicrobial formulations and methods for using the sameto effect both short term and persistent antimicrobial effects. Severalembodiments are particularly advantageous because they do not contributeto bacterial resistance to antibiotics.

Infections and Microorganisms

As discussed above, the potential spread of microorganisms and theassociated illnesses and infections are large. There are a variety ofdifferent types or microorganism-based infections or illnesses. A majorsource of illness is hospital acquired infections or health-careassociated infections. Once in a hospital, it is only a few hours untila patient's own microbial flora begin to acquire characteristics of thesurrounding pool of microorganisms. An infection that become clinicallyevident after 48 hours of hospitalization can be consideredhospital-acquired, while those developing after the patient isdischarged from the hospital can be considered healthcare-associated (ifthe organisms were acquired during the hospital stay). Use of theformulations disclosed herein, alone or in conjunction with infectionsurveillance programs can, in several embodiments, significantly reducehospital-acquired and healthcare-associated infections. Moreover, theiruse can also allow better prioritization of resources and efforts toimproving medical care.

Other environments present opportunities for spread of microorganismsand associated illnesses. Non-limiting examples include, dental offices,specialty surgical suites, physician offices, care-provided homes, orother medical care facilities, emergency vehicles (e.g., ambulances orfire engines), restaurants, food preparation areas (e.g., butcher shops,grocery stores), public or private transportation vehicles or venues,schools, playgrounds, sports or exercise venues, residential housing(e.g., dormitories, hostels, or hotels), etc. In several embodiments,the user (e.g., a non-medical professional) applies the formulation(s)to a surface susceptible to microorganism contamination (e.g., a usercan apply the formulation to his/her own skin).

As used herein, the term “microorganism” shall be given its ordinarymeaning and shall include, but not be limited to, viruses (including butnot limited to human immunodeficiency virus, herpes simplex virus,papilloma virus, parainfluenza virus, influenza, hepatitis, CoxsackieVirus, herpes zoster, measles, mumps, rubella, rabies, pneumonia,hemorrhagic viral fevers, H1N1, and the like), prions, parasites, fungi,mold, yeast and bacteria (both gram-positive and gram-negative)including, among others, Candida albicans, Aspergillus niger,Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), andStaphylococcus aureus (S. aureus), Group A streptococci, S. pneumoniae,Mycobacterium tuberculosis, Campylobacter jejuni, Salmonella, Shigella,and a variety of drug resistant bacteria. The terms microorganism andmicrobe shall be used interchangeably. Microbes can include wild-type,genetically-engineered or modified organisms. In several embodiments,the formulations and methods disclosed herein are for topical use ortreatment of a surface. However, in some embodiments, other uses ortreatments are used to achieve antimicrobial effects.

Antimicrobial Effects

The formulations and methods disclosed herein provide a variety ofantimicrobial effects. As used herein, the term “antimicrobial” shall begiven its ordinary meaning, and shall also refer to a substance compoundor formulation that kills or inhibits and/or otherwise reduces thenumber, growth or activity of microorganisms. It has been establishedthat hand washing is often incomplete, and that washing withnon-antimicrobial soap leaves no persistent effect. The formulationsprovided herein result in substantially greater reduction (both shortand long-term) of a target microorganism population than can be achievedthrough the use of plain soap. This translates to, in severalembodiments, a reduced risk of disease acquisition and/or organismtransmission. In several embodiments, the formulations reduce the riskof skin infection due to one's own resident skin flora and/or theacquisition of illness due to transmission of transient organisms fromoneself or from others via various transmission routes. In severalembodiments, the formulations are also used to facilitate wound healing(e.g., by reducing or preventing infection with one or moremicroorganisms). In several embodiments, the formulations are used totreat acne. In several embodiments, the formulations disclosed hereinmoisturize the skin in addition to reducing acne, but have limited sideeffects (e.g., are without drying effects on the skin). In severalembodiments, fungal infections can be treated and/or prevented byapplication of the formulations disclosed herein.

In some embodiments, the formulations kill the microorganisms (e.g.,they act as microbiocidal agents). In several embodiments, theformulations kill greater than 50% of the microorganisms contacted bythe formulation. In some embodiments, greater kill rates are achieved,for example, greater than 60%, greater than 70%, greater than 80%,greater than 85%, greater than 90%, greater than 91%, greater than 92%,greater than 93%, greater than 94%, greater than 95%, greater than 96%,greater than 97%, greater than 98%, greater than 99%, greater than99.9%, and overlapping ranges thereof. In several embodiments, theformulations provide a complete kill of the microorganisms (e.g., 100%).In several embodiments, a kill rate is achieved that substantiallyreduces the transmission or spread of the microorganism from person toperson (or person to surface, or surface to surface). In someembodiments, the number of microorganisms is substantially reduced aftercontact with the formulations as provided herein. For example, in someembodiments, a reduction in microorganism number of about 2 log isachieved. In other embodiments, greater log reductions are achieved, forexample, about 2.5 log reduction, about 2.8 log reduction, about 3.0 logreduction, about 3.2 log reduction, about 3.5 log reduction, about 4.0log reduction, about 5.0 log reduction, and overlapping ranges thereof.

In some embodiments, formulations and methods provided herein, inhibitthe microorganisms (e.g., they act as microbiostatic agents). In someembodiments, the inhibition is of the growth and/or reproduction,function, metabolism, and/or activity of the microorganisms. In someembodiments, the microorganisms are inhibited by 50% or more. In someembodiments, greater degrees of inhibition are achieved, for example,greater than 60%, greater than 70%, greater than 80%, greater than 85%,greater than 90%, greater than 91%, greater than 92%, greater than 93%,greater than 94%, greater than 95%, greater than 96%, greater than 97%,greater than 98%, greater than 99%, greater than 99.9%, and overlappingranges thereof. In several embodiments, the formulations provide acomplete kill of the microorganisms (e.g., 100%). In some embodiments,the inhibition is sufficient to cause the microorganism to no longer beinfectious; however, the microorganism is not necessarily dead. In otherembodiments, the inhibition (either directly or indirectly) leads to thedeath of the microorganism.

There exist a variety of antimicrobial formulations that kill onlycertain, limited, microbial populations. Thus, such formulations are notsuitable for killing a wide variety of microorganisms, but rather havelimited efficacy, and therefore limited suitability for applications inwhich a wide variety of microorganisms may be present. Moreover, somesuch formulations are less preferred because of various possible sideeffects, including but not limited to, drying of the skin, residuepost-application, skin irritation, skin rash development. These sideeffects, among others, can be exacerbated by frequent use, as may berequired in certain microorganism-rich environments.

The formulations disclosed herein, in several embodiments, function asbroad-spectrum antimicrobials. As such, the formulations disclosedherein kill or inhibit multiple different kinds of microorganisms, whenpresent, either alone or in combination with one another. Theformulations disclosed herein are particularly advantageous, in someembodiments, because they are effective against microorganisms that arerelatively easy to kill, as well as those organisms that are moredifficult to kill. The formulations disclosed herein are, in severalembodiments, effective at killing or inhibiting both gram-positive andgram-negative bacteria, drug-resistant bacteria, viruses, fungi, mold,yeasts, parasites, prions, as well as the other microorganisms disclosedherein, or combinations thereof. This broad-spectrum efficacy allowsfor, in some embodiments, a single formulation to be used to effectivelykill (or otherwise inhibit) these widely differing types ofmicroorganisms. As a result, transmission of these microorganisms (e.g.,from person-to-person or surface to surface) is reduced, therebyreducing or preventing spread of microorganisms. Existing formulations,in contrast, may only be effective against only one or just a few ofthese types of microorganisms. As a result, transmission of somemicroorganisms should be reduced and transmission of others would beunaffected. In certain microorganism rich environments, in particularthose with individuals susceptible to infection, such as hospitals orother health care facilities, existing formulations would thus be lessdesirable due to their lesser efficacy. In contrast, the formulationsdisclosed herein in several embodiments, with their broad-spectrumantimicrobial effects would provide substantially more benefit in suchenvironments do to the reduction and/or prevention of transmission ofthe wider spectrum of microorganisms.

Moreover, in several embodiments, the formulations disclosed hereinprovide a multiphase antimicrobial effect. As discussed herein, theformulations have both a short-term and a long-term (e.g., persistent)antimicrobial effect. The short-term effect, in several embodimentsprovides a rapid and robust inhibition or kill of microorganisms, whichtherefore results in a reduced possibility of transmission on ashort-term basis because of the substantial reduction in microorganismnumbers shortly after contact with the formulation. In some embodiments,the short-term effect occurs within a timeframe of several seconds up toabout several minutes. In several embodiments, the formulationsdisclosed herein provide extended antimicrobial effects, e.g., about 10minutes, about 15 minutes, about 25 minutes, about 35 minutes, andranges therebetween. In several embodiments, in addition to theshort-term and intermediate effects, the long-term (e.g., persistent)effects function to inhibit or kill microorganisms on a longertimescale, for example, on the order of hours.

In several embodiments, the timeframe in which killing or inhibition ofthe microorganism is achieved is a short-term timeframe. In severalembodiments, killing or inhibition of the microorganisms isinstantaneous (e.g., inhibition or death of the microorganism occurs assoon as the formulation context, the microorganism). In someembodiments, killing or inhibition occurs within about 2 seconds, withinabout 5 seconds, within about 7 seconds, within about 10 seconds, withinabout 15 seconds, within about 20 seconds, or within times in betweenthe timeframes listed above. For example, in some embodiments,substantial (or complete) inhibition or kill of microorganisms occurswithin about 10 to about 15 seconds, about 15 seconds to about 20seconds, about 20 seconds to about 30 seconds, about 30 seconds to about40 seconds, about 40 seconds to about 60 seconds, about 60 seconds toabout 90 seconds, about 90 seconds to about 120 seconds, and overlappingranges thereof. In some embodiments, the short-term effect persists forseveral minutes, for example, about 1 minute to about 2 minutes, about 2minutes to about 3 minutes, about 3 minutes to about 4 minutes, about 4minutes to about 5 minutes, about 5 minutes to about 15 minutes, about15 minutes about 30 minutes, about 35 minutes, about 40 minutes, about45 minutes, and overlapping ranges thereof. In several embodiments, theshort term killing of microorganisms is particularly advantageousbecause the microbial load is substantially or completely reduced priorto the point in time at which the microorganisms could spread. Inseveral embodiments, the rapid kill of microorganisms prevents thespread of the microorganisms which reduces the risk of transmission ofthe microorganism from a first subject to a second subject. Inconjunction with this effect, the rapid kill also reduces the riskand/or severity of infection that may be suffered by the first subjectas a result of the microorganisms (e.g., the fewer the microorganismspresent on the first subject, the less severe any resulting infectionis). Thus, not only do the formulations and methods disclosed hereinserve to reduce the spread of infectious microorganisms (which reducesthe number of individuals infected by the microorganisms) it also servesto reduce the impact of the remaining (if any) microorganisms on aparticular subject. In several embodiments, the rapid effect results inkill rates of greater than 90%, greater than 91%, greater than 92%,greater than 93%, greater than 94%, greater than 95%, greater than 96%,greater than 97%, greater than 98%, greater than 99%, greater than99.9%, and overlapping ranges thereof. In several embodiments, the rapideffect results in complete kill of the microorganisms (e.g., 100%).

For example, the formulations and methods disclosed herein provide apersistent effect resulting in substantial (or complete) inhibition orkill of microorganisms for timeframe ranging from, for example, about 20to about 40 minutes (e.g., about 35 minutes), about 40 to about 60minutes, about 60 to about 90 minutes, about 90 to about 120 minutes,about 120 to about 150 minutes, about 150 to about 180 minutes, about180 to about 240 minutes, and overlapping ranges thereof. In severalembodiments, the persistent effect of the formulations disclosed hereinresults for longer periods of time, ranging from about 30 min. to aboutone hour, about one hour to about two hours, about two hours to aboutthree hours, about three hours to about four hours, about four hours toabout six hours, about six hours to about eight hours, about eight hoursto about 12 hours, and overlapping ranges thereof. In some embodiments,certain formulations provide an even more extended persistent effect,such as from about four hours to about six hours, about six hours toabout eight hours, about eight hours to about 12 hours, and overlappingranges thereof. In several embodiments, the persistent effect achieveskill rates greater than 70%, greater than 80%, greater than 85%, greaterthan 90%, greater than 91%, greater than 92%, greater than 93%, greaterthan 94%, greater than 95%, greater than 96%, greater than 97%, greaterthan 98%, greater than 99%, greater than 99.9%, and overlapping rangesthereof. In several embodiments, the persistent effect complete kill ofthe microorganisms (e.g., 100%).

Advantageously, in several embodiments, the combined rapid effect andpersistent effect of the formulations disclosed herein reduces themicroorganism load in two important phases. First, the microbial load isreduced on a short-term basis, providing a significantly reduced degreeof initial spread of the microorganisms (e.g., from subject to subject,from subject to surface, etc.) and second, on a longer-term basis, whichfurther reduces the spread of the microorganisms, especially inconjunction with other sanitary practices (e.g., hand washing). Thequick initial kill is particularly advantageous because the formulationsactively kill the microorganisms prior to a time within which anindividual could engage in some other antimicrobial practice, forexample, washing hands. The persistent effect supplements the quickkill, and further reduces risk of later disease acquisition ortransmission of microorganisms.

Antimicrobial Formulations

There are provided for herein a variety of different types ofantimicrobial formulations in different forms. In several embodiments,the antimicrobial formulations are formulated as a hand wash or surgicalscrub. In several embodiments, such formulations are foaming. In severalembodiments, the foam generated is sufficiently thick to reduce run-offof the formulation during use. In several embodiments, the antimicrobialformulations are formulated as a soap (either bar soap or liquid/gelsoap). In several embodiments the antimicrobial formulations areformulated as a spray (e.g., an aerosol) or a gel. Non-gel sanitizersare provided in several embodiments. In several embodiments, theantimicrobial formulations are formulated as a lotion or other liquidform. In some embodiments, the formulations are provided as a cream orthickened lotion. In several embodiments, the formulations can beincorporated into other products, including but not limited to,cosmetics, body wash, shampoo, conditioner, sunscreens, insectrepellants, and the like.

In several embodiments, the formulations are formulated as, orincorporated into, anti-bacterial soap, liquid soap, bar soap, bathsoaps, cosmetic sunscreen preparations, deodorant for personal use, haircare preparations, hair conditioners, hair shampoo, hand lotions, handscrubs, hand soaps, non-medicated acne treatment preparations,non-medicated dental products and rinses (e.g., anti-plaque/anti-tartarformulations toothpaste, dental floss), acne treatment preparations,hand-sanitizing preparations, medicated dental products and rinses(e.g., anti-plaque/anti-tartar formulations, toothpaste, dental floss),medicated hair care preparations, medicated hand wash, medicated lotionsfor skin, medicated sunscreen, household deodorizer, cleaning agents andpreparations, cleaning preparations for cleaning surfaces, disinfectantsoaps, all-purpose disinfectants, anti-microbial handwash, antibacterialhand lotions, antibacterial spray, antimicrobial coatings to treat thegrowth of mold, mildew, bacteria and fungus on various surfaces,adhesive bandages, antiseptic wipes, antiseptics, bandages for skinwounds, bandages impregnated with antiseptic, first aid kits, fungalmedications, fungicides, gauze, germicides, gloves for medical use,surgical caps, surgical drapes, surgical gowns, surgical masks, surgicalscrub suits, surgical shoe covers, surgical devices and instruments,surgical sponges, sanitizing preparations for hospital use, sanitizingpreparations for household use, sanitizing preparations for use ininstitutional and industrial areas, sanitizing wipes, virucides and thelike.

In several embodiments the formulations comprise an alcohol. In severalembodiments, denatured alcohol is used. In several embodiments, thealcohol is about 200 proof, while in other embodiments, lesser proofalcohol is used (e.g., 40, 80, 100 proof). Alcohols include, but are notlimited to methanol, ethanol, isopropyl alcohol, butyl alcohol,pentanol, hexadecan-1-ol, ethane-1,2-diol, propane-1,2,3-triol,butane-1,2,3,4-tetraol, pentane-1,2,3,4,5-pentol,hexane-1,2,3,4,5,6-hexol, heptane-1,2,3,4,5,6,7-heptol, prop-2-ene-1-ol,3,7-dimethylocta-2,6-dien-1-ol, c3h3ohprop-2-in-1-ol,cyclohexane-1,2,3,4,5,6-hexol, and2-(2-propyl)-5-methyl-cyclohexane-1-ol. In several embodiments, thealcohol functions as an active killing agent, while in some embodiments,the alcohol is non-active. In several embodiments, the alcohol workssynergistically with one or more additional ingredients to provide theantimicrobial effects of the formulations. In several embodiments, thealcohol provides, at least in part, the rapid kill of microorganismsupon contact (or soon thereafter) with the microorganisms. In someembodiments, the alcohol also contributes, at least in part, to thepersistent effect of the formulations. Depending on the formulation, theamount of alcohol on a percent weight basis ranges from about 5% toabout 75%. In several embodiments, the amount of alcohol ranges fromabout 5% to about 7%, about 7% to about 10%, about 10% to about 12%,about 12% to about 14%, about 14% to about 16%, about 16% to about 20%,and overlapping ranges thereof. In some embodiments, the amount ofalcohol ranges from about 12% to about 16%, including about 12% to about13%, about 13% to about 13.5%, about 13.5% to about 14%, about 14% toabout 14.5%, about 14.5% to about 15%, bout 15% to about 15.5%, about15.5% to about 16%, and overlapping ranges thereof. In some embodiments,greater amounts of alcohol are used, for example, about 20% to about40%, about 40% to about 60% about 60% to about 65%, about 65% to about67%, about 67% to about 69%, about 69% to about 72%, about 72% to about75% and overlapping ranges thereof. In some embodiments, theformulations are alcohol free. In several embodiments, the formulationscomprise less than about 20% alcohol, e.g., less than about 15%, lessthan about 10%, less than about 5% or ranges therebetween.

In several embodiments, in addition to or in place of alcohol, chemicalagents are used to provide antimicrobial effects. In severalembodiments, chlorhexidine is used, and represents a non-limitingexample of a chemical antimicrobial agent. Chlorhexidine is particularlysuited for providing antibacterial effects, against both gram-positiveand gram-negative bacteria, though it provides additional antimicrobialeffects as well. In several embodiments, chlorhexidine, or otherchemical agent provides, at least in part, the immediate antimicrobialeffect of the formulation. In some embodiments, the chlorhexidine worksin conjunction with one or more additional component of the formulation,such as, for example, alcohol, to provide the immediate effect. In someembodiments, however, chlorhexidine also contributes, at least in part,to the persistent effect of certain formulations. Various salts ofchlorhexidine are used, depending on the embodiment. Non-limitingexamples include chlorhexidine digluconate, chlorhexidine diacetate,chlorhexidine phosphanilate, and chlorhexidine dihydrochloride.Combinations of the various salts, or the chlorhexidine base are used instill additional embodiments. In several embodiments, the amount ofchlorhexidine (or other chemical antimicrobial; on a % weight basis)ranges from about 0.01% to about 2%, including about 0.01% to about0.05%, about 0.05% to about 0.075%, about 0.075% to about 0.10%, about0.10% to about 0.15%, about 0.15% to about 0.175%, about 0.175%, toabout 0.19%, about 0.19% to about 0.20%, about 0.20% to about 0.22%,about 0.22% to about 0.25%, about 0.25% to about 0.30%, about 0.30% toabout 0.50%, about 0.50% to about 1.0%, about 1.0% to about 1.5%, about1.5% to about 2.0%, about 2.0% to about 3.0%, and overlapping rangesthereof. In certain formulations, surprisingly antimicrobial effects areachieved even though the amount of the chemical antimicrobial is belowthe “active kill” level as currently recognized by the FDA (e.g., 0.4for chlorhexidine). In some embodiments, the formulations are free ofchemical antimicrobials. For example, some embodiments of theformulations are chlorhexidine free.

In several embodiments, the formulations comprise one or more quaternaryammonium compounds. In some embodiments, the quaternary ammoniumcompounds are synthetic, while in other embodiments, they are naturallyoccurring. In some embodiments, synthetic quaternary ammonium compoundscomprising long alkyl chains are preferred, though those with shortalkyl chains are optionally used. In several embodiments, the quaternaryammonium compounds provide and/or supplement the persistent kill effectof the formulation. Non-limiting examples of quaternary ammoniumcompounds include benzalkonium chloride, benzethonium chloride,methylbenzethonium chloride, cetalkonium chloride, cetylpyridiniumchloride, cetrimonium, cetrimide, dofanium chloride, tetraethylammoniumbromide, didecyldimethylammonium chloride, and domiphen bromide, orcombinations of two or more thereof. In several embodiments, thequaternary ammonium compounds provide potent antimicrobial effects bydisrupting the cell membrane of microorganisms. In some embodiments,this disruption is sufficient to kill or inhibit the microorganism,while in some embodiments, the disruption provides a route by whichanother agent in the formulation provides the killing or inhibitingeffect. In several embodiments, the amount of quaternary ammoniumcompound (on a % weight basis) ranges from about 0.001% to about 2%,including about 0.001% to about 0.005%, about 0.005% to about 0.075%,about 0.075% to about 0.01%, about 0.01% to about 0.02%, about 0.02% toabout 0.03%, about 0.03% to about 0.04%, about 0.04% to about 0.05%,about 0.05% to about 0.075%, about 0.075% to about 0.10%, about 0.10% toabout 0.11%, about 0.11% to about 0.12%, about 0.12% to about 0.13%,about 0.13% to about 0.14%, about 0.14% to about 0.15%, about 0.15% toabout 0.175%, about 0.175% to about 0.19%, about 0.19% to about 0.20%,about 0.20% to about 0.22%, about 0.22% to about 0.25%, about 0.25% toabout 0.30%, about 0.30% to about 0.40%, about 0.40% to about 0.50%,about 0.50% to about 1.0%, about 1.0% to about 1.5%, about 1.5% to about2.0%, about 2.0% to about 3.0%, and overlapping ranges thereof. Inseveral embodiments, combinations of quaternary ammonium compounds areused. In certain such embodiments, the total amount of the quaternaryammonium compounds (on a per weight basis) is as described above. Inother embodiments, the amounts listed above are for each individualquaternary ammonium compound. In some embodiments, the formulations arefree of quaternary ammonium compounds or salts thereof.

In several embodiments, the antimicrobial efficacy of the quaternaryammonium compounds is pH dependent. Thus, in some embodiments, the pH ofthe formulation is adjusted to be within a certain range to improve thefunction of the quaternary ammonium compounds. For example, in someembodiments, the formulations range in pH from about 3.5 to about 4.0,about 4.0 to about 4.5, about 4.5 to about 5.5, about 5.5 to about 6.0,about 6.0 to about 6.5, about 6.0 to about 6.5, and overlapping rangesthereof. In several embodiments, the pH is more alkaline.

In several embodiments, certain ingredients in the formulationsupplement the antimicrobial efficacy of the formulation. In someembodiments, this supplementation is realized by making an activekilling agent more effective as an antimicrobial while in someembodiments, the additional ingredients also function as killing agents.In some embodiments, the supplementation is realized by virtue of adisinhibition of one or more other components of the formulation. Insome embodiments, the alternative killing ingredient supplements theoverall function of the formulation because of a different mechanism ofaction. In some embodiments, the formulation comprises a single activeingredient. In several embodiments, the presence of one or moreingredients potentiates the efficacy of one or more other ingredientseither through a synergistic mechanistic action, and/or by virtue ofreducing an inhibitory effect on another ingredient. For example,depending on the embodiment, certain components of the formulationsdisclosed herein are pH sensitive, in that their function is greaterwhen the formulation is within a certain pH range. In some embodiments,the additional components of the formulation are present to maintain thepH within that desired range, thereby improving the function of thefirst component.

In several embodiments, the formulation comprises one or more terpenesor derivatives thereof. In several embodiments, the terpenes arehemiterpenes (e.g., prenol or isovaleric acid), monoterpenes (e.g.,geraniol, limonene and terpineol), sesquiterpenes (e.g., farnesenes,farnesol), diterpenes (e.g., cafestol, kahweol, cembrene and taxadiene,sesterterpenes (e.g., geranylfarnesol), triterpenes (e.g., squaline,lanosterol, or cycloartenol), sesquarterpenes (e.g., ferrugicadiol andtetraprenylcurcumene), tetraterpenes (e.g., lycopene, carotenes), orpolyterpenes. Combinations of the various types of terpenes (orderivatives) may be used, in several embodiments. In some embodiments,the terpenes provide additional antimicrobial effects. In someembodiments, the terpenes contribute to the overall efficacy of theformulation by increasing the breadth of microorganisms that theformulation is effective against (e.g., increases in the kill spectrum).In some embodiments, the terpenes contribute to the persistent effect ofthe formulation. However, in some embodiments, the terpenes contributeto the rapid kill effect of the formulation. Additionally, in someembodiments, the terpenes function to buffer the pH of the formulation(by virtue of, at least in part, the anionic charge of the terpene). Insome embodiments, this buffering capacity aids in the stability of theformulation, or in some embodiments in the process of manufacturing theformulation. In still additional embodiments, the buffered pH is leadsto a reduced skin irritant potential of the formulation. In severalembodiments the buffered pH improves or potentiates the antimicrobialefficacy of one or more other components of the formulation. In severalembodiments, the anionic nature of the terpene allows buffering of othercations (in addition to or in place of H+, as discussed above) whichcontributes to the ionic stability of the formulation as a whole.

In several embodiments, the formulation comprises a terpene (orderivative thereof) in an amount ranging (on a % weight basis) betweenabout 0.1% to about 2%, including about 0.1% to about 0.2%, about 0.2%to about 0.3%, about 0.3% to about 0.4%, about 0.4% to about 0.45%,about 0.45% to about 0.5%, about 0.5% to about 0.55%, about 0.55% toabout 0.6%, about 0.6% to about 0.65%, about 0.65% to about 0.7%, about0.7% to about 1%, about 1% to about 1.5%, about 1.5% to about 2%, andoverlapping ranges thereof. In some embodiments, the formulations arefree of terpenes or derivatives thereof.

In addition, several embodiments of the formulation comprise vitamins oranalogs thereof. In several embodiments, the formulation comprisespanthenol, an alcohol analog of vitamin B5. In some embodiments,panthenol functions as an emollient and/or as a moisturizer. Asdiscussed above, in several embodiments, panthenol contributes to the pHof the formulation. In several embodiments, panthenol supplements theoverall antimicrobial efficacy of the formulation in a synergisticfashion. In several embodiments, panthenol is provided as a racemicmixture of D- and L-panthenol, while in some embodiments, theformulation comprises only one of the two enantiomers. In severalembodiments, the formulation comprises panthenol in an amount ranging(on a % weight basis) between about 0.1% to about 2%, including about0.1% to about 0.2%, about 0.2% to about 0.3%, about 0.3% to about 0.4%,about 0.4% to about 0.45%, about 0.45% to about 0.5%, about 0.5% toabout 0.55%, about 0.55% to about 0.6%, about 0.6% to about 0.65%, about0.65% to about 0.7%, about 0.7% to about 1%, about 1% to about 1.5%,about 1.5% to about 2%, and overlapping ranges thereof. In someembodiments, the formulations are panthenol free.

In several embodiments, the formulation comprises one or more biguanidepolymers, oligomers, or biguanides. For example, in some embodiments,the formulation comprises polyaminopropyl biguanide, which impartsbactericidal and/or fungicidal effects to the formulation (among othereffects, depending on the amount used in the formulation). In someembodiments, the polymer strands are disruptive to the integrity of themembranes of certain microorganisms, which leads to lethal effects forthe microorganisms. Additionally, polyaminopropyl biguanide, in severalembodiments, can induce lethal DNA damage in the microorganism.Advantageously, the greater degree of complexity of the cell membrane ofhigher species, such as humans, reduces the potential adverse effects ofthe polyaminopropyl biguanide on the cells of the higher species. Thus,polyaminopropyl biguanide, in several embodiments specifically targetsmicroorganisms. Advantageously, this preference for action on lesscomplex cell types also reduces the potential for dermal irritationresulting from use of the formulations. In some embodiments,polyaminopropyl biguanide is provided as a mixture of various lengthpolymers, and as such, the various lengths provide various efficaciesagainst certain types of microorganisms, increasing the spectrum ofpossible antimicrobial kills. In several embodiments, polyaminopropylbiguanide, or other biguanides, contribute, at least in part, to theimmediate antimicrobial effect of the formulations. In severalembodiments, polyaminopropyl biguanide, or other biguanides, contribute,at least in part, to the persistent antimicrobial effect of theformulations. In some embodiments, polyhexamethyl biguanide is used,either in addition to or in place of polyaminopropyl biguanide. In someembodiments, the formulations are free of biguanides. In severalembodiments, polyaminopropyl biguanide is particularly advantageousbecause it is limited in its inhibitory effects on other components ofthe formulation. In several embodiments, the amount of polyaminopropylbiguanide, or other biguanide, (on a % weight basis) ranges from about0.01% to about 3%, including about 0.01% to about 0.02%, about 0.02% toabout 0.03%, about 0.03% to about 0.04%, about 0.04% to about 0.05%,about 0.05% to about 0.075%, about 0.075% to about 0.080%, about 0.080%to about 0.090%, about 0.090% to about 0.10%, about 0.10% to about0.125%, about 0.125% to about 0.15%, about 0.15% to about 0.175%, about0.19% to about 0.20%, about 0.20% to about 0.22%, about 0.22% to about0.25%, about 0.25% to about 0.30%, about 0.30% to about 0.50%, about0.50% to about 1.0%, about 1.0% to about 1.2%, about 1.2% to about 1.3%,about 1.3% to about 1.4%, about 1.4% to about 1.5%, about 1.5% to about1.6%, about 1.6% to about 1.7%, about 1.7% to about 2.0%, about 2.0% toabout 3.0%, and overlapping ranges thereof.

Several embodiments of the formulations comprise additional agents.These agents may contribute, at least in part, to the antimicrobialeffect of the formulation (among other contributions). In someembodiments, the formulation comprises hydroxyethyl cellulose. In someembodiments, the hydroxyethyl cellulose comprises quaternizedhydroxyethyl cellulose (also known as polyquaternium-10). In someembodiments, the formulation comprises hydroxyethyl ethylcellulose inaddition to or in place of hydroxyethyl cellulose. In some embodiments,the formulations comprise a cellulose selected from the group consistingof calcium carboxymethyl cellulose, carboxymethyl cellulose acetatebutyrate, carboxymethyl hydroxyethylcellulose, cellulose acetate,cellulose acetate butyrate, cellulose gum, cellulose acetate propionate,cellulose acetate propionate carboxylate, cellulose succinate, cetylhydroxyethylcellulose, ethylcellulose, hydrolyzed cellulose gum,hydroxybutyl methylcellulose, hydroxyethylcellulose, hydroxyethylethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose,methylcellulose, hydroxypropyl methylcellulose acetate/succinate,methylcellulose, methyl ethylcellulose, methyl hydroxyethylcellulose,microcrystalline cellulose, potassium cellulose succinate, sodiumcellulose sulfate, and combinations thereof. In some embodiments, thehydroxyethyl ethylcellulose (or other cellulose) works synergisticallywith other ingredients to produce the persistent kill effect of theformulation. In some embodiments, this synergism results, at least inpart, from the limited inhibitory effect that the hydroxyethylethylcellulose has on other components of the formulation. In someembodiments, the hydroxyethyl ethylcellulose (or other cellulose) alsofunctions as a thickening agent, thereby changing the viscosity of theformulation to a desired level, depending on the embodiment. In severalsuch embodiments, the viscosity of the formulation contributes to itsspreadability (e.g., surface area coverage), which in turn can impactthe antimicrobial effect and/or use of the formulation. In severalembodiments, the amount of hydroxyethyl ethylcellulose (or othercellulose) or other cellulose (on a % weight basis) ranges from about0.001% to about 3%, including about 0.001% to about 0.005%, about 0.005%to about 0.0075%, about 0.075% to about 0.01%, about 0.01% to about0.02%, about 0.02% to about 0.03%, about 0.03% to about 0.05%, about0.05% to about 0.075%, about 0.075% to about 0.080%, about 0.080% toabout 0.090%, about 0.090% to about 0.10%, about 0.10% to about 0.125%,about 0.125% to about 0.15%, about 0.15% to about 0.175%, about 0.19% toabout 0.20%, about 0.20% to about 0.22%, about 0.22% to about 0.25%,about 0.25% to about 0.30%, about 0.30% to about 0.50%, about 0.50% toabout 1.0%, about 1.0% to about 1.5%, about 1.5% to about 2.0%, about2.0% to about 3.0%, and overlapping ranges thereof. In some embodiments,the formulations are free of one or all of cellulose, hydroxyethylethylcellulose, hydroxyethyl cellulose and/or quaternized hydroxyethylcellulose. In several embodiments, the formulation further compriselactic acid and/or lactate. Depending on the embodiment, the lactic acidand/or lactate is present in a total amount ranging from about 5% toabout 15% (by weight), including about 5% to about 7%, about 7% to about9%, about 9% to about 11%, about 11% to about 13%, about 13% to about15%, and overlapping ranges thereof.

Several embodiments of the formulations disclosed herein employ multipleagents with complementary function, achieved by different mechanisms ofaction. For example, in some embodiments, the formulations comprise oneor more zinc salts. The zinc salts can, in some embodiments, contributeto the immediate and/or persistent antimicrobial effect of theformulations and/or reduce the potential for dermal irritation resultingfrom use of the formulation. In some embodiments, zinc improves the feelof the formulation, e.g., the sensation of smoothness or silkiness whenthe formulation is used on a surface comprising skin and/or reduction inthe stickiness of the formulation. In several embodiments, zinc alsoreduces the drying time of certain formulations. In several embodiments,the drying time of the formulation ranges from about 5 to about 30seconds, including but not limited to about 5 to about 10 seconds, about10 to about 15 seconds, about 15 to about 20 seconds, about 20 to about25 seconds, about 25 to about 30 seconds, and overlapping rangesthereof. The zinc salts are water soluble in some embodiments, while inothers water insoluble zinc salts are used. Combinations of watersoluble and water insoluble may also be used, in certain embodiments.Non-limiting examples of zinc salts are zinc acetate, zinc butyrate,zinc gluconate, zinc glycerate, zinc glycolate, zinc formate, zinclactate, zinc picolinate, zinc propionate, zinc salicylate, zinctartrate, zinc undecylenate, zinc oxide, zinc stearate, zinc citrate,zinc phosphate, zinc carbonate, and zinc borate. In several embodimentsthe zinc is provided as a component of a zinc matrix. In someembodiments, one zinc salt is used, while in other embodiments, two ormore zinc salts are used. In several embodiments, the amount of zincsalt (on a % weight basis) ranges from about 0.01% to about 2%,including about 0.01% to about 0.05%, about 0.05% to about 0.075%, about0.075% to about 0.10%, about 0.10% to about 0.15%, about 0.15% to about0.175%, about 0.175% to about 0.19%, about 0.19% to about 0.20%, about0.20% to about 0.22%, about 0.22% to about 0.25%, about 0.25% to about0.30%, about 0.30% to about 0.50%, about 0.50% to about 1.0%, about 1.0%to about 1.5%, about 1.5% to about 2.0%, about 2.0% to about 3.0%, andoverlapping ranges thereof In some embodiments, such as those employingmore than one zinc salt, the total amount of all zinc salts (on a perweight basis) is as described above. Surprisingly, in some embodiments,lesser amounts of zinc in the formulation result in greater degrees ofantimicrobial activity. In other embodiments, the amounts listed aboveare for each individual zinc salt in the formulation. In someembodiments, the formulations are free of zinc or zinc-containingmatrices.

Several embodiments of the formulations disclosed herein employ one ormore moisturizing or emollient agents. As used herein, the terms“moisturizer” and “emollient” shall be given their ordinary meaning, andshall be used interchangeably, and shall refer to agents that functionto improve or maintain the hydration, softness, smoothness, and/orpliability of the skin and the like, in particular the epidermal layers.In some embodiments, the moisturizers function to help reduce dry skin,oily skin, aging skin (e.g., loss of elasticity), sensitive skin (e.g.,eczema). In some embodiments, combinations of moisturizers are used,with each member of the combination providing complementary function toimprove multiple aspects of the skin, for example, both dry andsensitive skin conditions are improved. In some embodiments, themoisturizing agents work as occlusives (e.g., they forming a thin filmon the surface of the skin to prevent any loss of moisture). In someembodiments, the moisturizing agents work as humectants (e.g., theyattract water from the air in order to moisturize the skin). In someembodiments, the moisturizing agents work to restore deficiencies in theskin (e.g., mineral, vitamins, lipids etc.). In some embodiments asingle moisturizing agent can work with multiple mechanisms of action.In several embodiments, the moisturizer compounds comprise one or moreof glycerin, dimethicone (including PEG-12 dimethicone). In severalembodiments, such agents also improve tactile feel of the formulation.

In several embodiments the moisturizers are present in the formulationin an amount (on a % weight basis) ranging from about 0.5% to about 10%.In several embodiments, depending on the embodiment, the moisturizersare present in the formulation in an amount ranging from about 0.5% toabout 1.0%, about 1.0% to about 1.1%, about 1.1% to about 1.2%, about1.2% to about 1.3%, about 1.3% to about 1.4%, about 1.4% to about 1.5%,about 1.5% to about 1.6%, about 1.6% to about 1.7%, about 1.7% to about1.8%, about 1.8% to about 1.9%, about 1.9% to about 2.0%, about 2.0% toabout 2.25%, about 2.25% to about 2.5%, about 2.5% to about 2.75%, about2.75% to about 3.0%, about 3.0% to about 3.25%, about 3.25% to about3.5%, about 3.5% to about 3.75%, about 3.75% to about 4.0%, about 4.0%to about 4.1%, about 4.1% to about 4.2%, about 4.2% to about 4.3%, about4.3% to about 4.4%, about 4.4% to about 4.5%, about 4.5% to about 4.6%,about 4.6% to about 4.7%, about 4.7% to about 4.8%, about 4.8% to about4.9%, about 4.9% to about 5.0%, about 5.0% to about 4.25%, about 5.25%to about 5.5%, about 5.5% to about 5.75%, about 5.75% to about 6.0%,about 6.0% to about 6.5%, about 6.5% to about 7.0%, about 7.0% to about7.5%, about 7.5% to about 8.0%, about 8.0% to about 8.5%, about 8.5% toabout 9.0%, about 9.0% to about 9.5%, about 9.5% to about 10.0%, andoverlapping ranges thereof. In several embodiments, greater amounts ofmoisturizers are present in the formulation, such as, for example, about10% to about 15%, about 15% to about 20%, about 20% to about 25%, about25% to about 30%, about 30% to about 50%, and overlapping rangesthereof. In several embodiments, the formulations are free ofmoisturizer or emollient compounds.

In several embodiments, the formulations further comprise one or morevitamins and/or minerals. For example, in several embodiments, theformulations include, but are not limited to, one or more of vitamin E,A, K, C, B, and the like. In several embodiments, vitamins incorporatedinto the formulation function as antioxidants and/or reduce toproduction or deleterious effects of free radicals. In some embodiments,the vitamins induce a more rapid cellular “turn over”, which results ina shorter time to bringing new cells to the surface of the skin, and asa result, a reduction in the rate of aging of the skin. In severalembodiments, the vitamins induce capillary (or other small vessel)vasoconstriction, which results in a reduction in skin discoloration orother skin blemishes. In several embodiments, vitamins reduce theappearance of age spots and/or reduce discrepancies in the tone ofvarious parts of the skin (e.g., result in a more uniform skin color).In some embodiments, vitamins also reduce the adverse skin effectsassociated with acne (e.g., skin reddening, scarring, discoloration,etc.). In several embodiments, the formulations are free of vitamins. Inseveral embodiments, antimicrobial formulations for conditions such asacne or inflammatory skin conditions comprise an exfoliant. Exfoliantsmay be included in hand soaps and surgical scrubs. In some embodiments,formulations comprise a penetrant to enhance contact of the ingredientsinto the layers or pores of the skin.

Additional ingredients, depending on the embodiment include, but are notlimited to, citric acid; foaming agents such as amine oxide surfactants(e.g., lauramine oxide); viscosity builders/foam stabilizers includingbut not limited to, lauramine oxide, cocamidopropyl betaine, disodiumlaureth sulfosuccinate, cocamidopropylamine oxide, lauramidopropylbetaine, lauramide DEA, methyl hydroxyethylcellulose (also known asStructure® CEL), stearamidopropyl dimethylamine); additional emollientagents such as, for example cetrimonium chloride, hydroxyethyl cocamide(which can also enable the fragrances to be solubilized, other oils andactives in certain formulation embodiments); compounds to maintainemulsions (e.g., prevent emulsion separation) such as, cetyl alcoholand/or cetearyl alcohol; preservative agents, including but not limitedto, for example, methychloroisothiazolinone, methylisothiazolinone (alsoknown as Microcare® ITL), euxyl k 700 (benzyl alcohol, phenoxyethanol,potassium sorbate, and tocopherol) and the like; plant extracts, such asaloe extract, essential oils (including but not limited to volatileoil(s) obtained from a plant or an animal source comprising an activeagent which may be, for example but not by way of limitation, amonoterpene or sesquiterpene hydrocarbon, alcohol, ester, ether,aldehyde, ketone, or oxide. Examples of these essential oils include,but are not limited to, almond oil, ylang-ylang oil, neroli oil,sandalwood oil, frankincense oil, peppermint oil, lavender oil, jasmineabsolute, geranium oil bourbon, spearmint oil, clove oil, lemongrassoil, cedarwood oil, balsam oils, and tangerine oil, or agents from theessential oils including but not limited to 1-citronellol,alpha-amylcinnamaldehyde, lyral, geraniol, farnesol, hydroxycitronellal,isoeugenol, eugenol, eucalyptus oil and eucalyptol, lemon oil, linalool,and citral) cocoa butter, and the like; fragrances including, but notlimited to, floral, citrus, fruit, nut, vegetable, botanicals, woods,resin, mint, musk, spices, and the like; dimethicone; glycerine;conditioning agents/emulsifiers, such as Incroquat® Behnyl TMS (cetearylalcohol, benhentrimonium methosulfate). Depending on the embodiments,such additional ingredients may be added alone or in combination, and invarious amounts ranging from about 0.005% to about 10% (by weight),including about 0.005% to about 0.0075%, about 0.0075% to about 0.01%,about 0.01% to about 0.015%, about 0.015% to about 0.02%, about 0.02% toabout 0.04%, about 0.04% to about 0.06%, about 0.06% to about 0.08%,about 0.08% to about 0.10%, about 0.10% to about 0.20%, about 0.20% toabout 0.30%, about 0.30% to about 0.40%, about 0.40% to about 0.50%,about 0.50% to about 0.60%, about 0.60% to about 0.70%, about 0.70% toabout 0.80%, about 0.80% to about 0.90%, about 0.90% to about 1.0%,about 1.0% to about 1.1%, about 1.1% to about 1.2%, about 1.2% to about1.3%, about 1.3% to about 1.4%, about 1.4% to about 1.5%, about 1.5% toabout 1.6%, about 1.6% to about 1.7%, about 1.7% to about 1.8%, about1.8% to about 1.9%, about 1.9% to about 2.0%, about 2.0% to about 2.25%,about 2.25% to about 2.5%, about 2.5% to about 2.75% about 2.75% toabout 3.0% about 3.0% to about 3.25% about 3.25% to about 3.5% about3.5% to about 3.75% about 3.75% to about 4.0% about 4.0% to about 4.25%about 4.25% to about 4.5% about 4.5% to about 4.75% about 4.75% to about5.0% about 5.0% to about 5.5% about 5.5% to about 6.0% about 6.0% toabout 6.5%, about 6.5% to about 7.0%, about 7.0% to about 7.5%, about7.5% to about 8.0%, about 8.0% to about 8.5%, about 8.5% to about 9.0%,about 9.0% to about 9.5%, about 9.5% to about 10%, and overlappingranges thereof. Some embodiments are free one or more of theseadditional ingredients.

Several embodiments of the formulations also comprise water (optionallydeionized and/or distilled), which, depending on the embodiment, may beused to adjust the viscosity and/or feel of the formulation, solubilizeone or more components of the formulation, adjust the pH, and/orfinalize the concentrations of the formulation (bring the formulation toits final volume). Thus, water, in some embodiments is added in a rangefrom about 1% to about 99% of the formulation, including from about 1%to about 10%, about 10% to about 20%, about 20% to about 30%, about 30%to about 40%, about 40% to about 50%, about 50% to about 60%, about 60%to about 70%, about 70% to about 80%, about 80% to about 90%, about 90%to about 99%, and overlapping ranges thereof.

In several embodiments, the formulation comprises at least onequaternary ammonium salt selected from the group consisting ofbenzethonium chloride and benzalkonium chloride. In several embodiments,the formulation comprises a terpene or derivative thereof and at leastone quaternary ammonium salt selected from the group consisting ofbenzethonium chloride and benzalkonium chloride. Optionally, theformulations can further include one or more of zinc (e.g., a zincsalt), a chemical antimicrobial agent (e.g., chlorhexidine digluconate),a biguanide polymer (e.g., polyaminopropyl biguanide), a moisturizingagent (e.g., dimethicone), a viscosity builder/thickener (e.g.,hydroxyethyl ethylcellulose), spreading agent (e.g., glycerine), aconditioning agent (e.g., a cationic conditioning agent such asincroquat), and/or alcohol. These formulations exhibit antimicrobialactivity against one or more of the following microorganisms, viruses(including but not limited to human immunodeficiency virus, herpessimplex virus, papilloma virus, parainfluenza virus, influenza,hepatitis, Coxsackie Virus, herpes zoster, measles, mumps, rubella,rabies, pneumonia, hemorrhagic viral fevers, H1N1, and the like),prions, parasites, fungi, mold, yeast and bacteria (both gram-positiveand gram-negative) including, among others, Candida albicans,Aspergillus niger, Escherichia coli (E. coli), Pseudomonas aeruginosa(P. aeruginosa), and Staphylococcus aureus (S. aureus), Group Astreptococci, S. pneumoniae, Mycobacterium tuberculosis, Campylobacterjejuni, Salmonella, Shigella, and a variety of drug resistant bacteria.Antimicrobial activity can be achieved on various surfaces, such as skinor inorganic surfaces (e.g., countertops, door handles, faucets,telephones beds/bed frames, bed linens, medical equipment, computers,writing instruments, surgical equipment, etc.). The formulation may bein gel, cream, lotion, powder or spray form, and may be embedded withina material or coated onto a material. Surgical garments and gloves maycontain (e.g., be treated with) several embodiments of the formulationsdescribed herein. Formulations described herein may also be used intransportation, tourist and food industries. Preservation of food andbeverages may be accomplished by using several embodiments of theformulations described herein.

Depending on the particular embodiment, certain ingredients listedherein that are generally associated with a particular function may alsohave other functions, including functions within other generalcategories listed, as well as other functions that are appreciated inthe art.

Adjunct Beneficial Effects

In addition to the benefits of the broad-spectrum efficacy of theformulations disclosed herein and the advantages of the short andlong-term effects, formulations disclosed herein provide otherbeneficial effects as well. For example, in several embodiments, the useof moisturizers or other emollient compounds provide a substantialmoisturizing effect to the skin of the user of the formulation. Not onlydoes this improve the general tactile feel of the subject's skin, theincreased moisture in the skin provides a more effective natural barrierto prevent microbial infections. For example, properly moisturized skinis less prone to cracking and flaking or other forms of irritation thatmay foster microbial growth. In conjunction, the formulations disclosedherein are formulated in a manner that reduces the drying effect of theskin. Thus, in certain embodiments, the components of the formulationswork synergistically with one another to reduce the ability ofmicroorganisms to populate the skin of an individual into improve theoverall health of the skin of an individual. In some embodiments, theformulations are applied to broken or damaged skin (e.g., a laceration,scrape, cut, burn, incision, etc.), and thereby reduce or preventmicrobial growth, which concurrently reduces the potential for infectionand/or scarring.

In view of these benefits beyond the anti-microbial activity of theformulations, the formulations and several embodiments foster betteruser compliance than existing antimicrobial formulations. This isbecause, at least in part, the formulations disclosed herein provide abetter tactile feel, less residue, or an improvement in skin health as aresult of their use. This increased user compliance subsequently reducestransmission of microorganisms by virtue of the fact that theformulations of are used more regularly. In several embodiments,however, that “regular” use does not necessarily, depending on theembodiment, correspond to increased frequency of use. This is because,at least in part, the persistent effect of the formulations disclosedherein allows a longer duration of time between uses, yet still allowsfor substantial (or complete) inhibition or kill microorganisms.Advantageously, the persistent effect and the resultant long-terminhibition or kill of microorganisms in some embodiments, reduces theoverall cost to a facility for supplying an antimicrobial formulationbecause the required frequency of use is reduced. In some embodiments,however, more frequent use occurs, such as, for example, in certainsituations where microorganism load is anticipated to be greater thannormal. Alternatively, in some embodiments, users of the formulation canself-select an appropriate frequency of use, based on their ownenvironments and or desires with respect to the tactile feel of theformulation, and the resultant moisturizing qualities.

In some embodiments, the formulations are prepared without the use ofcertain art-recognized antimicrobial compounds. For example, in severalembodiments, the formulations are triclosan-free. In some embodiments,the formulations are free from silver (or other metal) nanoparticles. Inseveral embodiments, the formulations are free of nonorganic antibioticsand/or organic biocides.

In several embodiments, an additional added benefit of the formulationsis the shelf-life stability. The formulations are suitable for storageafter production for a period ranging from several months up to severalyears. For example, in several embodiments the formulations are stable(e.g., retain antimicrobial activity) for about 6 months to about 12months, about 12 months to about 18 months, about 18 months to about 24months, or overlapping ranges thereof. Additionally in some embodiments,the formulations are heat stable. This is advantageous, in severalembodiments b/c the formulations can be shipped to, stored, and used inrural environments where the temperatures may be higher (e.g.,facilities without air conditioning). In several embodiments, theformulations are non-flammable. In several embodiments, the formulationsare non-combustible.

Moreover, the formulations are useful in a variety of differentdispensers or containers. The formulations, depending on the embodiment,are suitable for dispensing from a wall mount dispenser, a hand pumpcontainer, a tube or bottle, an aerosol container, or a tub.

In several embodiments, the above-referenced compounds can be used invarious, non-limiting combinations in order to provide a formulationthat is optimized for a particular purpose (e.g., targetingpreferentially a particular type (or types) of microorganism, killing orinhibiting at a certain level, etc.). Moreover, in several embodiments,the above referenced compounds that make up any given embodiment of theformulation may recognized by those of skill in the art by commercial ortrade names, chemical nomenclature of formula, or the InternationalNomenclature for Cosmetic Ingredients designation.

EXAMPLES

The following studies were designed to test several embodiments of theformulations and methods disclosed herein in terms of their ability toreduce microbial load over time. Various microbes were tested in theexperiments, and antimicrobial effects were measured at various timepoints to assess immediate and persistent antimicrobial effects.

Several examples discussed in more detail below involve antimicrobialtesting using Staphylococcus aureus (or MRSA, where specificallyindicated) and/or E. coli. These bacteria represent two examples, onegram positive, and one gram negative, respectively, of a variety ofmicroorganisms that are responsible for illness and disease throughoutvarious environments/communities and, given the relative severity ofinfection with these (or other) microorganisms, reduction in thecolonization of surfaces with such microorganisms can, in severalembodiments, significantly reduce spread and incidence of infectionscaused by these bacteria, other drug resistant bacteria, and othervarieties of microorganisms.

Example 1 Testing of Persistent and Non-Persistent Soaps

Duration testing was carried out using a modification of the ASTM 1882method for hand soap. In brief, agar plates were grown to a populationof 5×10³ S. aureus and then treated with one of several formulations.Microbial counts were taken a 2 minutes, 60 minutes, and 240 minutes.After treatment, the percent reduction and log reduction were calculatedfor each time point.

TABLE 1 Percentage reduction in S. aureus post-treatment. % Reductionfor Each Formulation Formulation 1 Formulation 2 Formulation 3 TimePoint (contains (Persistent; (Non-persistent; (minutes) triclosan)triclosan free) triclosan free) 2 100 99.67 91.64 60 100 92.90 58.82 24099.96 83.53 46.55

TABLE 2 Log₁₀ reduction in S. aureus post-treatment. % Reduction forEach Formulation Formulation 1 Formulation 2 Formulation 3 Time Point(contains (Persistent; (Non-persistent; (minutes) triclosan) triclosanfree) triclosan free) 2 3.50 1.49 1.09 60 3.69 1.16 0.40 240 3.50 0.790.27

These results demonstrate that each formulation resulted in substantialand/or complete reduction in microbial load, depending on the time pointtested and the formulation. Formulations 2 and 3, persistent andnon-persistent formulations, respectively, provide an unexpected degreeof efficacy against S. aureus given that the formulations aretriclosan-free. In several embodiments, other formulations are alsotriclosan-free, which advantageously allows efficacious antimicrobialeffects without the adverse effects associated with triclosan.

In several embodiments, the persistent and non-persistent soapformulations are particularly advantageous due to the rate at which theantimicrobial effects are realized. For example, in several embodiments,the antimicrobial effects (e.g., substantial or complete kill) areachieved after a fraction of the time required for antimicrobial effectsto be achieved with other existing formulations. In several embodiments,the time to achieve antimicrobial effects ranges from about 2 to about 5seconds of hand washing with the formulation, about 5 to about 7 secondsof hand washing, about 7 to about 10 seconds of hand washing, about 10to about 12 seconds of hand washing, about 12 to about 15 seconds ofhand washing, about 15 to about 18 seconds of hand washing, about 18 toabout 20 seconds of hand washing, about 20 to about 25 seconds of handwashing_(s) about 25 to about 30 seconds of hand washing, andoverlapping ranges thereof. This rapid effect is advantageous becauseexisting formulations require a significantly longer time of constanthand washing, on the order of minutes. Because the vast majority ofusers of existing formulations fail to perform the hand washing for therequisite period of time, those users are not receiving the fullantimicrobial effect that such formulations provide. Thus, the presentlydescribed formulations compensate, in several embodiments, for a degreeof user error by inducing a more substantial antimicrobial effectdespite the lesser degree of time of actual hand washing.

Example 2 Testing of Variant Persistent and Non-Persistent SoapFormulations

Duration testing was carried as described above using a modification ofthe ASTM 1882 method for hand soap. Variants of certain persistent andnon-persistent soap formulations were tested. Percent reduction and logreduction were calculated for each time point.

TABLE 3 Percentage reduction in S. aureus post-treatment. % Reductionfor Each Formulation Formulation Time Formulation 2 with Formulation 2Point 2 with 0.7% 5% Benzyl with 2.5% (minutes) Formulation 2 FarnesolAlcohol DMSO 2 97.6 97.6 93.8 96.6 60 97.8 94.9 88.4 85.7 240 95.4 91.490.9 86.7

TABLE 4 Log₁₀ reduction in S. aureus post-treatment. Log Reduction forEach Formulation Formulation Time Formulation 2 with Formulation 2 Point2 with 0.7% 5% Benzyl with 2.5% (minutes) Formulation 2 Farnesol AlcoholDMSO 2 1.63 1.63 1.2 1.47 60 1.37 1.30 0.94 0.85 240 1.34 1.06 1.04 0.88

These data appear to demonstrate that, certain formulations provideunexpectedly efficacious antimicrobial activity, despite using lesserquantities of certain agents. Addition of either benzyl alcohol or DMSO(which would be expected to increase the antimicrobial effects of theformulations) also failed to increase the antimicrobial effect over thatachieved by Formulation 2 alone, in one embodiment. In severalembodiments the various components of the formulations disclosed hereinhave been optimized to provide antimicrobial effects that are unexpectedin view of conventional wisdom that suggests that “more is better”. Inseveral embodiments, as discussed above other essential oils (e.g.,either in addition to or in place of farnesol) can be used and,depending on the embodiment different amounts can be used. In severalembodiments, the use of other essential oils, either alone or incombination with farnesol continue to provide potent antimicrobialeffects, even when less of that particular component is used, ascompared to what conventional wisdom indicates and/or other commercialformulations employ. The formulations disclosed herein comprise, inseveral embodiments, agents that synergistically work in combination toprovide surprisingly efficacious immediate and/or persistentantimicrobial effects.

Example 3 Antimicrobial Testing of Lotion Formulations

Duration of efficacy was carried out using a modification of the ASTM E1882 method. In brief, the test employed pig skin samples (approximately16 cm²) as the test application surface for the formulations. The pigskin was exposed to an inoculum of 5×10³ Escherichia coli and thentreated with one of a variety of formulations. Bacterial counts weremade at 2 minutes, 60 minutes, and 240 minutes.

TABLE 5 Percentage reduction in E. Coli after leave-on treatment TimePoint (minutes) Formulation Description 2 60 240 A Base Lotion 94 91 87(comprises farnesol and benzethonium chloride) B Base + 0.6% zincgluconate 94 92 88 C Base + 0.3% zinc gluconate + 94 92 85 0.3% zinclactate D Base + 0.1% zinc gluconate + 97 96 89 0.1% zinc lactate EBase + 1.5% polyaminopropyl 98 96 95 biguanide F Base + 0.1% EucalyptusOil 95 94 91 G Base + 0.25% Quaternized 95 91 87 hydroxyethyl celluloseH Base + 0.1% Barcleans 95 94 81 I Base + 0.3% zinc gluconate + 93 92 810.3% zinc lactate + 0.1% eucalyptus oil + 1.5% polyaminopropyl biguanideJ Base + 0.25% Quaternized 96 91 86 hydroxyethyl cellulose + 0.3% zincgluconate + 0.3% zinc lactate + 0.1% Barcleans K Formulation J + 2.5%DMSO 97 94 88

TABLE 6 Log reduction in E. Coli after leave-on treatment Time Point(minutes) Formulation Description 2 60 240 A Base Lotion 1.22 1.08 0.91(comprises farnesol and benzethonium chloride) B Base + 0.6% zincgluconate 1.21 1.13 0.93 C Base + 0.3% zinc gluconate + 1.19 1.13 0.820.3% zinc lactate D Base + 0.1% zinc gluconate + 1.54 1.38 0.97 0.1%zinc lactate E Base + 1.5% polyaminopropyl 1.82 1.37 1.34 biguanide FBase + 0.1% Eucalyptus Oil 1.30 1.21 1.06 G Base + 0.25% Quaternized1.36 1.09 0.91 hydroxyethyl cellulose H Base + 0.1% Barcleans 1.29 1.260.73 I Base + 0.3% zinc gluconate + 1.16 1.13 0.72 0.3% zinc lactate +0.1% eucalyptus oil + 1.5% polyaminopropyl biguanide J Base + 0.25%Quaternized 1.56 1.07 0.86 hydroxyethyl cellulose + 0.3% zincgluconate + 0.3% zinc lactate + 0.1% Barcleans K Formulation J + 2.5%DMSO 1.55 1.26 0.95

As shown in the data above, several embodiments of the formulationsdisclosed herein are effective at reducing microorganism load over anextended period of time. In several embodiments, the persistent effectkills at least about 80% of the microorganisms for a period of time ofat least 4 hours. In some embodiments, greater percentages ofmicroorganism death are achieved, for example from about 80% to about85%, about 85% to about 90%, about 90% to about 91%, about 91% to about92%, about 92% to about 93%, about 93% to about 94%, about 94% to about95%, about 95% to about 96%, about 96% to about 97%, about 97% to about98%, about 98% to about 99%, about 99% to about 100%, and overlappingranges thereof. In several embodiments, the persistent effect lasts forgreater than 4 hours, e.g., about 4 to about 5 hours, about 5 to about 6hours, about 6 to about 7 hours, about 7 to about 8 hours, about 8 toabout 9 hours, about 9 to about 10 hours, or overlapping ranges thereof.In several embodiments, the persistent effect is beneficial in that thenumber of uses a subject requires is reduced over a given period oftime. This not only makes the formulations more cost-effective, but alsoincreases user compliance, as compliance is less (or non-) intrusiveinto an activity or activities performed by a subject. In someembodiments, however, there are also benefits to more frequent uses(e.g., improved skin quality and feel due to the formulations).

The results presented above illustrate the surprisingly effective natureof certain formulations as broad spectrum antimicrobials having atwo-phase antimicrobial effect (e.g., a short-term very high kill phaseand a persistent high kill phase). For example, the formulationcomprising the base lotion plus 1.5% polyaminopropyl biguanide, with a98% reduction at 2 minutes and 96% reduction at 60 minutes, stillmaintains a 95% reduction in E. coli. (1.134 log reduction), even at 240minutes. The unexpectedly effective dual action immediate and persistenteffects are, in several embodiments, a result of the synergisticinteractions between the components of the formulation. Similarly, thebase lotion supplemented with eucalyptus oil, a non-limiting example ofan essential oil, shows not only a high immediate microbial reduction,but a substantially maintained persistent effect as well. In someembodiments, the formulations can comprise only the immediate effects.

The range of applications of the formulations disclosed herein, and thecorresponding methods, is wide. For example the formulations can beemployed in a variety of environments, including but not limited tohospitals, dental offices, specialty surgical suites, physician offices,care-provided homes, or other medical care facilities, emergencyvehicles (e.g., ambulances or fire engines), restaurants, foodpreparation areas (e.g., butcher shops, grocery stores), public orprivate transportation vehicles or venues, schools, playgrounds, sportsor exercise venues, residential housing (e.g., dormitories, hostels, orhotels), etc.

Example 4 Antimicrobial Testing of Sanitizer Formulations Against E.Coli

Testing was performed as described above, and surfaces were treated withvarious sanitizer formulations. Bacterial counts were made a 2 minutes,60 minutes, and 240 minutes.

TABLE 7 Percentage Reduction in E. Coli After Sanitizer Application TimePoint (minutes) Formulation Description 2 60 240 L CommercialFormulation 1 96 96 76 M Sanitizer Formulation 1 98 96 91 N SanitizerFormulation 2 95 94 88

TABLE 8 Log Reduction in E. Coli After Sanitizer Application Time Point(minutes) Formulation Description 2 60 240 L Commercial Formulation 11.52 1.37 0.63 M Sanitizer Formulation 1 1.84 1.46 1.03 N SanitizerFormulation 2 1.28 1.21 0.94

The results presented above illustrate the surprisingly effective natureof the sanitizer formulations disclosed herein as broad spectrumantimicrobials having a two-phase antimicrobial effect (e.g., ashort-term very high kill phase and a persistent high kill phase). Asshown above, application of sanitizer formulation 1 results in a 2%improvement in immediate microbial reduction at 2 minutes (98%reduction; 1.84 log), which is maintained at a surprisingly high level,even out to 240 minutes. At that long-term test point, the commercialformulation resulted in only 76% microbial reduction, while sanitizerformulation 1 maintained a 91% reduction (1.03 log). This dual-actioneffect, based in several embodiments on the synergistic nature of thecomponents of the sanitizer formulation, is particularly advantageous asit is not only reduces microbial load on contact with a surface, butprevents transmission of the microbes over a long period of time. Inseveral embodiments, these effects are also achieved, at least in part,due to the physical characteristics of the formulation. For example, inseveral embodiments the formulations have a viscosity that allows forimproved coverage of a surface (e.g., spreadability) without beingoverly liquid (which in some cases could lead to undesired flow off ofthe target surface). In some embodiments, the formulations also have arapid drying time, which prevents, at least to some degree, theunintended loss of the formulation from a target surface. In severalembodiments, low absorption of the formulation into the surface (e.g.,skin) also improves the coverage. In yet other embodiments, theformulation is configured to absorb into or adhere to the surface of theskin to achieve persistence.

These results demonstrate that persistence can be achieved with avariety of formulations. However, in several embodiments certainformulations provide unexpectedly greater degrees of antimicrobialactivity over time, even with high levels of short-term efficacy. Asdiscussed above, the combination of short and long-term efficacy, aswell as the wide spectrum effects (e.g., as demonstrated by the abilityto kill the more difficult gram-negative bacteria) yield formulationswhich are unexpectedly beneficial in view of existing formulations.

Example 5 Sanitizing Formulations Demonstrate Broad Spectrum and RapidAntimicrobial Effects

In recent years, alcohol-based hand sanitizers developed into thedominant products for hand antisepsis in the absence of water. Productshaving between about 60% and about 95% alcohol are currently the onlysanitizers recommended for use in hospitals by the CDC (see, e.g.,Centers for Disease Control and Prevention, Hand Hygiene in HealthcareSettings, May 19, 2011, http://www.cdc.gov/handhygiene/). In suchsettings, the alcohol concentration functions to rapidly kill of a broadspectrum of bacteria, including gram positive and negative bacteria.

However, certain alcohol sanitizers still present significantlimitations and drawbacks to their use. Studies have shown that usingcertain high concentration alcohol sanitizers in hospitals is equivalentto handwashing alone, but does not result in the desired reduction inthe occurrence of hospital-acquired infections (HAIs), which can impactabout 1 in 20 hospitalized patients and kill nearly 90,000 Americanseach year. Certain high concentration alcohol sanitizers can result in asignificant drying effect on the hands which can reduce compliance(e.g., reduced and/or ineffective frequency of use). For example,alcohol products can dry the hands with repeated use may result incracked and painful skin (which could lead to conditions that promoteinfections). Even trained healthcare workers may only follow recommendedhand hygiene protocols a portion the time, which can be, at least inpart, due to the drying nature of alcohol-based products. Reduceduse/compliance is clinically relevant because alcohol products areeffective until such time that they evaporate, typically about 15seconds. After this time, hands may become immediately re-contaminated,as there is no persistent activity (another limitation of alcohol-basedproducts). Further, the reduced compliance, and increased likelihood ofcontamination can foster germ transmission from patients and staff, andvice versa.

Another significant limitation of certain alcohol sanitizers is therelatively lower efficacy against many viruses, for example, theylargely ineffective against non-enveloped viruses. Non-enveloped virusesare a significant cause of disease in healthcare settings and in homes.Non-limiting of non-enveloped viruses include, but are not limited to,Norovirus (leading cause of acute gastroenteritis or the “stomach flu”),rhinoviruses (common colds), rotavirus (severe diarrhea, especially inchildren), adenoviruses (tonsillitis and conjunctivitis) and HepatitisA, among others. The relative lack of efficacy of alcohol against suchviruses clinical significance, as a study involving use of an alcoholsanitizer found the alcohol sanitizer to be less effective than rinsingwith water alone against Norovirus, only reducing concentration by0.14-0.34 log. An additional study reported that nursing homesfacilities that relied on alcohol products instead of handwashing weremore than six times as likely to have a Norovirus outbreak. Further,cruise ships, another closed environment where Norovirus is dangerous,have averaged 16 outbreaks a year, despite the widespread use of alcoholsanitizers. In elementary school settings, where germ transmission canrapidly lead to widespread illness-related absenteeism, studies showedthat alcohol-based products, decreased illness-related absenteeism byonly 19% in one study and result in no change in a another. However,antiseptic lotion and antiseptic foaming soap formulations according toseveral embodiments herein were provided to two schools for placement incommonly used areas (e.g., by the door, in the bathrooms). Throughcollection of over 6,000 data points, these formulation reducedabsenteeism by nearly 42% among students (41.6%) and nearly 25% amongteachers and staff (24.7%). These results demonstrate that theformulations disclosed herein, in several embodiments, outperformalcohol-based sanitizers and can coordinately reduce absenteeism, lostwork/study time, etc. These non-laboratory setting data show that theformulations herein have a real world efficacy, which was furtherinvestigated in the laboratory controlled example discussed below.

The present experiments were designed to evaluate the rapid andbroad-spectrum efficacy of alcohol-based and non-alcohol-basedsanitizing products according to the compositions disclosed herein thatseek to address the limitations of certain traditional alcohol-basedproducts.

Methods

The experiments compared an alcohol-based Antiseptic (comprising,according to one embodiment, 70% ethyl alcohol v/v, “EtOH Antiseptic”),a water-based Antiseptic Lotion (comprising a quaternary ammonium, suchas, according to one embodiment, benzethonium chloride at 0.2%,“Antiseptic Lotion”), and a Foaming Hand Soap (comprising, according toone embodiment, benzethonium chloride, 0.2%, “Foaming Soap”). This studyused an in vitro Time-Kill protocol to evaluate the antisepticcompositions when challenged with 25 different microorganism species asdescribed in the FDA Tentative Final Monograph (see FDA Tentative FinalMonograph, Topical Antimicrobial Products for Over-the-Counter Use, 21CFR 333 and 369, Federal Register 59; 116, 1994). Each product wasevaluated at a 99% concentration, and the percent and log reductionswere determined following exposure times of 15, 30, and 60 seconds.Agar-plating was performed in duplicate. A combination of clinicalisolates and lab strains were used in the testing.

Neutralization studies of each product were also performed versusBacteroides fragilis, Escherichia coli, Staphylococcus aureus aureus,and Streptococcus pneumoniae to ensure the neutralizing solutionemployed (Butterfield's Phosphate Buffer solution with productneutralizers) was effective and non-toxic to each of the representativechallenge species.

Results

The log reduction (and percent) due to the EtOH Antiseptic and theAntiseptic Lotion are shown in Table 9.

TABLE 9 Log and Percent Reduction of Various Microorganisms Over TimeEtOH Antiseptic Foaming Organism Time Antiseptic Lotion SoapAcinetobacter 15 sec >6.57 >6.57 >6.57 baumannii (99.9999%) (99.9999%)(99.9999%) (ATCC #19606) 30 sec >6.57 >6.57 >6.57 (99.9999%) (99.9999%)(99.9999%) 60 sec >6.57 >6.57 >6.57 (99.9999%) (99.9999%) (99.9999%)Bacteroides fragilis 15 sec >7.22 3.87 >7.22 (ATCC# 25285) (99.9999%)(99.9864%) (99.9999%) 30 sec >7.22 6.92 >7.22 (99.9999%) (99.9999%)(99.9999%) 60 sec >7.22 3.92 >7.22 (99.9999%) (99.988%) (99.9999%)Candida albicans 15 sec >6.16 2.17 0.0460 (ATCC# 10231) (99.9999%)(99.3218%) (10.0346%) 30 sec >6.16 2.75 0.1305 (99.9999%) (99.8208%)(25.9516%) 60 sec >6.16 3.65 0.5067 (99.9999%) (99.9777%) (68.8581%)Candida tropicalis 15 sec >6.14 4.14 0.8468 (ATCC #750) (99.9999%)(99.9927%) (85.7706%) 30 sec >6.14 >6.14 1.3251 (99.9999%) (99.9999%)(95.2688%) 60 sec >6.14 >6.14 2.2873 (99.9999%) (99.9999%) (99.4839%)Enterobacter 15 sec >6.27 >6.27 >6.27 aerogenes (99.9999%) (99.9999%)(99.9999%) (ATCC #13048) 30 sec >6.27 >6.27 >6.27 (99.9999%) (99.9999%)(99.9999%) 60 sec >6.27 >6.27 >6.27 (99.9999%) (99.9999%) (99.9999%)Enterococcus 15 sec >6.20 5.38 >5.20 faecalis (99.9999%) (99.9996%)(99.9994%) (ATCC# 29212) 30 sec >6.20 >6.20 >5.20 (99.9999%) (99.9999%)(99.9994%) 60 sec >6.20 >6.20 >5.20 (99.9999%) (99.9999%) (99.9994%)Enterococcus 15 sec >5.90 >5.90 3.4638 faecium (99.9999%) (99.9999%)(99.9656%) (ATCC #6057) 30 sec >5.90 >5.90 4.6021 (99.9999%) (99.9999%)(99.9975%) 60 sec >5.90 >5.90 >4.94 (99.9999%) (99.9999%) (99.9988%)Escherichia coli 15 sec >6.14 >6.14 3.2318 (ATCC #11229) (99.9999%)(99.9999%) (99.9414%) 30 sec >6.14 >6.14 3.8865 (99.9999%) (99.9999%)(99.9870%) 60 sec >6.14 >6.14 4.9389 (99.9999%) (99.9999%) (99.9988%)Escherichia coli 15 sec >5.27 >5.27 >5.28 (ATCC #25922) (99.9995%)(99.9995%) (99.9995%) 30 sec >5.27 >5.27 >5.28 (99.9995%) (99.9995%)(99.9995%) 60 sec >5.27 >5.27 >5.28 (99.9995%) (99.9995%) (99.9995%)Haemophilus 15 sec >6.23 >6.23 >5.23 influenzae (99.9999%) (99.9999%)(99.9994%) (ATCC #33930) 30 sec >6.23 >6.23 >5.23 (99.9999%) (99.9999%)(99.9994%) 60 sec >6.23 >6.23 >5.23 (99.9999%) (99.9999%) (99.9994%)Klebsiella oxytoca 15 sec >6.17 >6.17 >6.17 (ATCC #13182) (99.9999%)(99.9999%) (99.9999%) 30 sec >6.17 >6.17 >6.17 (99.9999%) (99.9999%)(99.9999%) 60 sec >6.17 >6.17 >6.17 (99.9999%) (99.9999%) (99.9999%)Klebsiella 15 sec >6.17 >6.17 2.5875 pneumoniae (99.9999%) (99.9999%)(99.7415%) pneumoniae (ATCC #4352) 30 sec >6.17 >6.17 >6.17 (99.9999%)(99.9999%) (99.9999%) 60 sec >6.17 >6.17 >6.17 (99.9999%) (99.9999%)(99.9999%) Micrococcus luteus 15 sec >5.40 >5.40 >4.40 (ATCC #7468)(99.9996%) (99.9996%) (99.9960%) 30 sec >5.40 >5.40 >4.40 (99.9996%)(99.9996%) (99.9960%) 60 sec >5.40 >5.40 >4.40 (99.9996%) (99.9996%)(99.9960%) Proteus mirabilis 15 sec >6.08 >6.08 3.0634 (ATCC #7002)(99.9999%) (99.9999%) (99.9136%) 30 sec >6.08 >6.08 4.5048 (99.9999%)(99.9999%) (99.9969%) 60 sec >6.08 >6.08 >6.08 (99.9999%) (99.9999%)(99.9999%) Pseudomonas 15 sec >6.29 >6.29 4.1264 aeruginosa (99.9999%)(99.9999%) (99.9925%) (ATCC #15442) 30 sec >6.29 >6.29 4.5095 (99.9999%)(99.9999%) (99.9969%) 60 sec >6.29 >6.29 5.0090 (99.9999%) (99.9999%)(99.9990%) Pseudomonas 15 sec >5.92 >5.92 4.9675 aeruginosa (99.9999%)(99.9999%) (99.9989%) (ATCC #27853) 30 sec >5.92 >5.92 >5.92 (99.9999%)(99.9999%) (99.9999%) 60 sec >5.92 >5.92 >5.92 (99.9999%) (99.9999%)(99.9999%) Serratia marcescens 15 sec >6.48 5.63 0.0424 (ATCC #14756)(99.9999%) (99.9998%) (9.3023%) 30 sec >6.48 >6.48 0.1637 (99.9999%)(99.9999%) (31.3953%) 60 sec >6.48 >6.48 0.1564 (99.9999%) (99.9999%)(30.2326%) Staphylococcus 15 sec >6.11 4.09 0.6469 aureus (99.9999%)(99.9919%) (77.4517%) aureus (ATCC #6538) 30 sec >6.11 >6.11 1.2928(99.9999%) (99.9999%) (94.9035%) 60 sec >6.11 >6.11 2.8102 (99.9999%)(99.9999%) (99.8452%) Staphylococcus 15 sec >5.56 >5.56 0.8706 aureus(99.9997%) (99.9997%) (86.5278%) aureus (ATCC #29212) 30 sec >5.56 >5.561.1447 (99.9997%) (99.9997%) (92.8333%) 60 sec >5.56 >5.56 1.4407(99.9997%) (99.9997%) (96.3750%) Staphylococcus 15 sec >5.45 >5.45 >4.45epidermidis (99.9996%) (99.9996%) (99.9965%) (ATCC #12228) 30sec >5.45 >5.45 >4.45 (99.9996%) (99.9996%) (99.9965%) 60sec >5.45 >5.45 >4.45 (99.9996%) (99.9996%) (99.9965%) Staphylococcus 15sec >5.05 >5.05 >4.05 haemolyticus (99.9991%) (99.9991%) (99.9910%)(ATCC #29970) 30 sec >5.05 >5.05 >4.05 (99.9991%) (99.9991%) (99.9910%)60 sec >5.05 >5.05 >4.05 (99.9991%) (99.9991%) (99.9910%) Staphylococcus15 sec >4.95 >4.95 >3.95 hominis (99.9989%) (99.9989%) (99.9888%)hominis (ATCC #27844) 30 sec >4.95 >4.95 >3.95 (99.9989%) (99.9989%)(99.9888%) 60 sec >4.95 >4.95 >3.95 (99.9989%) (99.9989%) (99.9888%)Staphylococcus 15 sec >5.88 >5.88 >4.88 saprophyticus (99.9999%)(99.9999%) (99.9987%) (ATCC #35552) 30 sec >5.88 >5.88 >4.88 (99.9999%)(99.9999%) (99.9987%) 60 sec >5.88 >5.88 >4.88 (99.9999%) (99.9999%)(99.9987%) Streptococcus 15 sec >6.29 >6.29 >5.29 pneumoniae (ATCC(99.9999%) (99.9999%) (99.9995%) #6303) 30 sec >6.29 3.05 >5.29(99.9999%) (99.9100%) (99.9995%) 60 sec >6.29 4.93 >5.29 (99.9999%)(99.9988%) (99.9995%) Streptococcus 15 sec >6.11 >6.11 >5.11 pyogenes(99.9999%) (99.9999%) (99.9992%) (ATCC #19615) 30 sec >6.11 >6.11 >5.11(99.9999%) (99.9999%) (99.9992%) 60 sec >6.11 >6.11 >5.11 (99.9999%)(99.9999%) (99.9992%)

The EtOH Antiseptic eliminated at least 99.999% of all organisms testedwithin 15 seconds. With respect to the duration of time to achieve thatamount of microbial reduction, substantially all of the antimicrobialeffects occurred in the first 15 seconds, as no differences weredetected in kill percentage between the 15 second and 60 secondtimepoints.

The Antiseptic Lotion was also highly effective against the testedmicroorganisms, destroying more than 99.99% of 24 of 25 organisms within15 seconds. The Antiseptic Lotion showed a slightly reduced efficacyagainst Candida albicans, a fungal yeast, showing a 99.32% kill oncontact. Against the 23 bacterial strains, the Antiseptic Lotionaveraged a 99.999% kill within the first 15 seconds. In the analysis ofduration of time to achieve the kill, 99.97% of the antimicrobial effectwas achieved within the first 15 seconds of contact of the AntisepticLotion with the microorganisms. The results of the duration analysis areshow in Table 10.

TABLE 10 Duration of Time to Achieve Maximum Antimicrobial Effects EtOHAntiseptic Antiseptic Lotion Difference Average Kill at 15 seconds99.9998% 99.9715% 0.028% Average Kill at 30 seconds 99.9998% 99.8990%0.011% Average Kill at 60 seconds 99.9998% 99.9988% 0.001% Percent oftotal effect achieved in    100%  99.97%  0.03% first 15 seconds

The Foaming Soap had a less rapid effect against certain microorganisms,buy still exhibited substantial anti-microbial effects against nearlyall the microorganisms tested. The longer time to efficacy, in severalembodiments, is due to the formulation of the soap and the intended useof certain embodiments of the formulation (e.g., hand washing forrecommended times and with recommended temperature water). Despitehaving lesser antimicrobial effects on an acute (e.g., 15-30 seconds) ascompared to the EtOH Antiseptic and the Antiseptic Lotion, the FoamingSoap still exhibited microbial reduction of 99% or greater at 60 secondsagainst 22 of the 25 microorganisms tested. The rate of antimicrobialeffects is particularly surprising and unexpected, as the antisepticlotion, in several embodiments, is alcohol-free. Thus, in severalembodiments, the antiseptic lotion provides rapid effects (and longerterm effects which are discussed more below) without the potentialadverse effects associated with certain alcohol based products. Thus, inseveral embodiments, the Foaming Soap is particularly advantageous as anantimicrobial agent, particularly when handwashing conditions of 60seconds or more are employed. In several embodiments, color changingagents or other indicators are provided to indicate optimal efficacy(e.g., the soap changes color or consistency after 60 seconds).

The experiments in this Example employ a standard protocol, and as such,the results can be compared against products that contain variousconcentrations of ethanol (or other alcohol) alone, to compare theimmediate kill of the antiseptic products disclosed herein against astandard alcohol product. As shown in Table 11, a commercially availablealcohol-based sanitizer (61% alcohol) was tested against analcohol-based antiseptic according to several embodiments disclosedherein, and an antiseptic lotion according to several embodimentsdisclosed herein.

TABLE 11 Comparison of Antimicrobial Activity Against CommerciallyAvailable Alcohol-Based Sanitizer AVAGARD D EtOH Antiseptic SanitizerAntiseptic Lotion S. aureus 99.1% 99.9997% 99.9997% S. epidermis >99.9%99.9996% 99.9996% K. pneumoniae >99.9% 99.9999% 99.9999% P.aeruginosa >99.9% 99.9999% 99.9999% E. coli >99.9% 99.9999% 99.9999% S.pneumoniae >99.9% 99.9999% 99.9999% S. pyogenes 98.0% 99.9999% 99.9999%S. marcescens >99.9% 99.9999% 99.9998% E. faecalis >99.9% 99.9999%99.9996%

These results demonstrate that both the EtOH Antiseptic according toseveral embodiments disclosed herein and the Antiseptic Lotion accordingto several embodiments disclosed herein achieved not only a rapid killeffect but also a broad-spectrum kill against a wide variety ofmicroorganisms. The antimicrobial effect of the EtOH Antiseptic and theAntiseptic Lotion according to several embodiments disclosed herein wasgreater than that achieved by a commercially available Sanitizer(AVAGUARD D), reducing bacterial load by over ˜2 log against themicroorganisms tested (notably, these include both E. coli and S.aureus).

Surprisingly, the immediate, broad-spectrum kill profile issubstantially similar between the EtOH Antiseptic and the AntisepticLotion (which is water-based). Results of this time-kill testingeliminate the concern that a non-alcohol product cannot have the samerapid antimicrobial benefits as an alcohol-based sanitizer. Furthermore,this time-kill study demonstrated that both the Antiseptic andAntiseptic Lotion had a greater immediate kill than a traditional,alcohol-only sanitizer. In several embodiments, the EtOH sanitizer,despite its relatively high alcohol concentration, is not associatedwith several of the adverse effects of traditional alcohol-basedsanitizers, due to, for example, additional ingredients that providemoisturizing and/or protective effects on the skin. Moreover theseresults indicate that the Antiseptic Lotion (which is water based) is atleast equivalent to certain alcohol-based products currently used inhealthcare facilities for immediate, broad-spectrum kill. These superiorresults are particularly surprising given that the Antiseptic Lotion iswater-based, given that alcohol is responsible for a substantial amountof antimicrobial effects in many alcohol-based products. In severalembodiments, the use of a water-based product may reduce skinirritation, improve compliance, eliminate concerns about flammabilityand storage, and add a persistent effect to the products, each of whichcontributes to the increased efficacy of the Antiseptic Lotion. Inseveral embodiments, the formulations are non-flammable. In severalembodiments, the formulations are non-combustible.

Example 6 Efficacy of Sanitizing Formulations in In Vivo HealthcarePersonnel Wash Testing

As discussed above, Hospital Acquired Infections (HAI) are a majorsource of morbidity and mortality. Health-care associated pathogenstransmitted not only from infected or draining wounds, but also fromfrequently colonized areas of normal, intact patient skin. For example,patients with diabetes, patients undergoing dialysis for chronic renalfailure, and those with chronic dermatitis are likely to have areas ofintact skin that are colonized with S. aureus. While data suggests thatdirect patient contact and respiratory-tract care are most likely tocontaminate the fingers of caregivers and duration of patient-careactivity was strongly associated with the intensity of bacterialcontamination of health care provider hand, patient gowns, bed linen,bedside furniture, and other objects in the patient's immediateenvironment can easily become contaminated with patient flora.Transmission from patient to health care workers can then result inre-transmission to another patient, another health care worker, or toindividuals outside the health care setting. In any event, thetransmission and re-transmission of microorganisms can result inwide-spread infections and associated illnesses.

Efforts to reduce the infection of health care workers and spread ofmicroorganisms (and incidence if HAIs) are commonly manifest in handwashprocedures. The present experiment was designed to evaluate the efficacyof certain sanitizing formulations disclosed herein when used accordingto 21 CFR parts 333 and 369, Topical Antimicrobial Drug Products forOver-the-Counter Human Use; Tentative Final Monograph (TFM) forHealth-Care Antiseptic Drug Products; Proposed Rule.

Methods

A total of fifty-two subjects were subjected to eleven consecutive handcontaminations with Serratia marcescens (ATCC #14756). Subjects wereassigned randomly to use of one of the two test products or thereference product (e.g., each subject was assigned to use one, and onlyone, of the three test materials). After the first contamination, abaseline sample was obtained, and thereafter, the remaining 10contaminations were performed, each following by application of one ofthe test products. Microbial samples were taken after productapplications 1, 3, 7, and 10. Data from a total of forty-six subjectswere included in the statistical analysis with data from 16 subjectsincluded for Test Product #1, data from 15 subjects included for TestProduct #2, and data from 15 subjects included for the ReferenceProduct. Mean log₁₀ reductions of S. marcescens were used to evaluatethe antimicrobial efficacy of each test product. The two test productswere the EtOH Antiseptic and the Antiseptic Lotion (from Example 5). Thecommercially available reference product was HIBICLENS®, which comprisesa 4% (w/v) solution of chlorhexidine gluconate.

A stock culture of S. marcescens was prepared by asepticallytransferring a lyophilized pellet to approximately 5.0 mL of sterileTryptic Soy Broth (TSB), which was then incubated at 25° C.±2° C. for 24hours±4 hours. A 2-L flask containing approximately 1,000 mL TSB wasinoculated with 1.0 mL of the 24-hour broth culture, and incubated for24 hours±4 hours at 25° C.±2° C. Prior to any withdrawal of culture,whether for hand contamination or for numbers assay, the suspension wasstirred or swirled. The suspension was assayed for number of organismsat the beginning and end of the use period. A suspension was not used intesting for more than 8 hours.

Each subject was in testing for 3.5 to 4.5 hours on a single day.Subjects' fingernails clipped were to a free edge of ˜1 mm. All jewelrywas removed from the hands and arms prior to the inception of the studyperiod. Using tap water, a practice hand-inoculation procedure wasperformed by each subject, to reduce subject-to-subject variation do toapplication procedures. A 5.0-mL aliquot of a suspension containingapproximately 1.0×10⁹ CFU/mL of S. marcescens was transferred into eachsubject's cupped hands in three successive volumes of approximately 1.5mL, 1.5 mL, and 2 mL, respectively. The inoculum then was distributedevenly over both hands, not reaching above the wrists, via continuousmassage for 45 seconds.

After a timed 2-minute air-dry, the Glove Juice Sampling Procedure wasperformed. That procedure involved the placement of powder-free, sterilelatex gloves were on the subjects' hands and instillation of 75.0 mLStripping Suspending Fluid (SSF) into each of the gloves. The wristswere secured, and attendants massaged the hands through the gloves in astandardized manner for 60 seconds. Thereafter, a sample from each glovewas taken to establish baseline recovery values. Additionally, a 5.0-mLaliquot of the glove juice was removed from each of the gloves andseparately diluted in 5.0 mL Butterfield's Phosphate Buffer Solutionwith product neutralizers (BBP++) (dilution 10⁰). The 10⁰ dilutions thenwere serially diluted in BBP++, as appropriate. Following the GloveJuice Sampling Procedure was a 30-second handwash using nonmedicatedsoap and a 30-second rinse.

Subsequent contaminations were performed by distributing an additional5.0 mL of the S. marcescens suspension over both hands, not reachingabove the wrists, via gentle continuous massage for 45 seconds. A timed2-minute air-dry was performed, followed by the application of randomlyassigned test material to each subject according to the following.

For the EtOH Antiseptic, an amount sufficient to thoroughly wet thesubjects' hands, approximately 2 mL to 3 mL, was dispensed intosubjects' cupped dry hands. Subjects rubbed the test product over theentire surface of the hands and fingers paying special attention tonails and nail beds, and continued to rub into the skin until dry. Priorto any sample, subjects waited an additional 5 minutes after productappeared dry.

For the Antiseptic Lotion, an amount sufficient to thoroughly wet thesubjects' hands, approximately 1 mL to 2 mL, was dispensed intosubjects' cupped dry hands. Subjects rubbed the test product over theentire surface of the hands and fingers paying special attention tonails and nail beds, and continued to rub into the skin until dry. Priorto any sample, subjects waited an additional 5 minutes after productappeared dry.

For the Reference Product, subjects wet their hands and forearms and 5mL of the Reference Product was dispensed into the subjects' cuppedhands. Subjects then rubbed their hands together and lathered theproduct onto their hands and forearms for 30 seconds, followed by arinse of their hands for 30 seconds. For washes that were followed by asample, the hands were gloved wet. Subjects lightly dried hands with adisposable paper towel after washes not followed by a sample.

Each subject completed the above contamination/product application atotal of 10 consecutive times, with a minimum of 5 minutes betweenmicrobe/product applications. Following contamination/productapplication cycle 9 the subjects assigned to use one of the two testproducts rinsed their hands for 30 seconds. The hands were sampled forresidual S. marcescens after contamination/product application cycles 1,3, 7, and 10. All samples were performed using the Glove Juice SamplingProcedure (discussed above).

Samples were plated on duplicate spiral plates prepared from appropriatedilutions using Tryptic Soy Agar with product neutralizers (TSA+). Theplates were incubated at 25° C.±2° C. for approximately 48 hours, oruntil sufficient growth was observed. S. marcescens produces redcolonies, and only those colonies were counted. Colonies were countedand data recorded using the computerized Q-COUNT™ plate-counting system.If 10⁰ spiral plates gave an average count of zero, the average platecount was expressed as 1.00×10¹. A neutralization study (data not shown)assured that the neutralizers used in the recovery medium effectivelyquenched the antimicrobial activity of the test materials and were nottoxic to the challenge species. Study procedures were based on ASTM E1054-08, Standard Test Methods for Evaluation of Inactivators ofAntimicrobial Agents.

The estimated Log₁₀ number of viable microorganisms recovered from eachsubject's hand was designated the “R-value,” the adjusted average log₁₀colony count measurement from each sample. Each R-value was determinedusing the following formula: R=log₁₀ [75×C_(i)×10^(−D)×2], where 75 isthe amount (mL) of stripping solution instilled into each glove, C_(i)is the arithmetic average colony count of the two plate counts from eachsample at a particular dilution level, D is the dilution factor, and 2is the neutralization dilution.

Log₁₀ reductions from baseline populations recovered from each of asubject's hands were calculated by subtracting the log₁₀ number ofviable microorganisms recovered from the hands following productapplication from the log₁₀ number of viable microorganisms recovered forbaseline. Mean and standard deviation data were generated on the log₁₀data from baseline samples, post-product application samples, and thereductions from baseline. These data are shown in Tables 12-16.

TABLE 12 Statistical Data of the Mean Log₁₀ Microbial Recoveries of S.marcescens and Mean Log₁₀ Microbial Reductions from Baseline FollowingSubjects' Use of EtOH Antiseptic Sample Minimum Maximum Sample Size MeanValue Value Baseline Log₁₀ Microbial 32 9.14 7.86 9.47 RecoveryPost-Product Application 1 32 6.32 5.42 7.15 Log₁₀ Microbial RecoveryPost-Product Application 3 32 6.72 4.89 8.23 Log₁₀ Microbial RecoveryPost-Product Application 7 32 5.53 3.18 7.32 Log₁₀ Microbial RecoveryPost-Product Application 10 32 5.43 3.18 6.42 Log₁₀ Microbial RecoveryPost-Product Application 1 32 2.82 2.13 3.42 Log₁₀ Reduction fromBaseline Post-Product Application 3 32 2.42 0.97 3.95 Log₁₀ Reductionfrom Baseline Post-Product Application 7 32 3.62 1.75 5.83 Log₁₀Reduction from Baseline Post-Product Application 10 32 3.72 2.64 5.59Log₁₀ Reduction from Baseline

TABLE 13 Statistical Data of the Mean Log₁₀ Microbial Recoveries of S.marcescens and Mean Log₁₀ Microbial Reductions from Baseline FollowingSubjects' Use of Antiseptic Lotion Sample Minimum Maximum Sample SizeMean Value Value Baseline Log₁₀ Microbial 30 9.26 8.96 9.44 RecoveryPost-Product Application 1 30 7.62 6.79 8.43 Log₁₀ Microbial RecoveryPost-Product Application 3 30 7.64 5.94 8.64 Log₁₀ Microbial RecoveryPost-Product Application 7 30 6.54 4.33 8.29 Log₁₀ Microbial RecoveryPost-Product Application 10 30 6.15 4.09 8.17 Log₁₀ Microbial RecoveryPost-Product Application 1 30 1.64 0.86 2.50 Log₁₀ Reduction fromBaseline Post-Product Application 3 30 1.63 0.66 3.37 Log₁₀ Reductionfrom Baseline Post-Product Application 7 30 2.73 0.99 4.95 Log₁₀Reduction from Baseline Post-Product Application 10 30 3.11 1.12 5.10Log₁₀ Reduction from Baseline

TABLE 14 Statistical Data of the Mean Log₁₀ Microbial Recoveries of S.marcescens and Mean Log₁₀ Microbial Reductions from Baseline FollowingSubjects' Use of HIBICLENS ® Sample Minimum Maximum Sample Size MeanValue Value Baseline Log₁₀ Microbial 30 9.14 8.35 9.42 RecoveryPost-Product Application 1 30 6.28 5.42 7.01 Log₁₀ Microbial RecoveryPost-Product Application 3 30 5.53 4.68 6.20 Log₁₀ Microbial RecoveryPost-Product Application 7 30 4.91 3.96 5.67 Log₁₀ Microbial RecoveryPost-Product Application 10 30 4.72 3.66 5.57 Log₁₀ Microbial RecoveryPost-Product Application 1 30 2.85 1.59 3.76 Log₁₀ Reduction fromBaseline Post-Product Application 3 30 3.61 2.32 4.34 Log₁₀ Reductionfrom Baseline Post-Product Application 7 30 4.23 2.78 5.06 Log₁₀Reduction from Baseline Post-Product Application 10 30 4.42 2.78 5.42Log₁₀ Reduction from Baseline

To ensure that the reductions detected were due to product applicationand not to falling microbial population numbers, the populations of theS. marcescens inoculum suspensions used in testing were over the testingperiod and are presented in Table 15.

TABLE 15 Initial and Final Populations of the S. marcescens InoculumSuspension Initial Population Final Population Test Date Group (CFU/mL)(CFU/mL) Apr. 4, 2013 1 (PM) 1.42 × 10⁹ 1.69 × 10⁹ Apr. 5, 2013 2 (PM)1.78 × 10⁹ 1.65 × 10⁹ Apr. 12, 2013 3 (AM) 1.62 × 10⁹ 1.62 × 10⁹ Apr.12, 2013 4 (PM) 1.87 × 10⁹ 1.69 × 10⁹ Apr. 16, 2013 5 (AM) 1.57 × 10⁹1.62 × 10⁹ Apr. 16, 2013 6 (AM) 1.59 × 10⁹ 1.77 × 10⁹ Apr. 16, 2013 7(PM) 1.69 × 10⁹ 1.50 × 10⁹

Summary statistics were also generated on the log₁₀ reductions frombaseline for the EtOH Antiseptic, the Antiseptic Lotion, and forHIBICLENS® (Mölnlycke Health Care, AB), following product applications 1and 10, and are presented in Table 16.

TABLE 16 Statistical Summary Log₁₀ Microbial Reductions from Baseline95% Std. Dev. N Mean Min Max Std. Dev. Conf. Int. EtOH Antiseptic 322.8213 2.1313 3.4204 0.3409 0.254295-0.50429 Post-Application 1 EtOHAntiseptic 32 3.715 2.641 5.591 0.685 0.511085-1.10354 Post-Application10 Antiseptic Lotion 30 1.6417 0.8550 2.4967 0.4352 0.321774-0.65379Post-Application 1 Antiseptic Lotion 30 3.107 1.117 5.099 0.9310.688543-1.39899 Post-Application 10 HIBICLENS ® 30 2.8527 1.5889 3.75910.4983 0.368502-0.74873 Post-Application 1 HIBICLENS ® 30 4.419 2.7755.420 0.574 0.424471-0.86245 Post-Application 10

A two-way analysis of variance (ANOVA) was performed to compare theefficacy of the EtOH Antiseptic, the Antiseptic Lotion, and HIBICLENS®.The ANOVA results (calculations not shown) are depicted graphically inFIGS. 1 and 2. These data show that HIBICLENS® and the EtOH Antisepticshowed the same log reduction (from baseline) at application #1. TheEtOH Antiseptic showed a 3.5 log reduction, even after 10re-inoculations with S. marcescens. While the Antiseptic Lotion showedlower reduction at Application 1 as compared to the other productstested, after 10 re-inoculations with S. marcescens, the AntisepticLotion showed greater than a 3.0 log reduction in microorganism load.

These data demonstrate that, the EtOH antiseptic passed the 2.0 log₁₀reduction at Application 1 and the 3.0 log₁₀ reduction at Application10, per the FDA Healthcare Personnel Handwash Requirements. While theAntiseptic Lotion, at least according to this specific example, did notpass the 2.0 log₁₀ reduction at Application 1, it passed the 3.0 log₁₀reduction at Application 10 (see FIG. 3). Of note is that the protocolperformed in this example was designed specifically for wash-off handproducts (e.g., soaps intended to be rinsed from the hands after use).This protocol employs a significant initial load of microorganism, inthe view that the rinsing of the hands after application will result insome (potentially substantial) loss of microorganism. Thus, the higherinitial load is to ensure that meaningful microorganism data can beobtained (e.g., lower initial load with hand washing could lead tominimal microorganism counts at a first time point, providingpotentially misleading results into the efficacy of a formulation). Inthe example discussed above, the antiseptic lotion was not washed off(in accordance with the intended use of one embodiment of theformulation), though the FDA protocol initial microorganism load wasemployed. As such, the reduction in the microorganism count is solelyattributable to the application of the antiseptic formulation (and notbased on any wash off of the product or microorganism). Thus, theefficacy of the Antiseptic Lotion, while not meeting the initialApplication 1 FDA threshold numerically, is significant, in that thethreshold was nearly reached, without the “benefit” of a rinse removingmicroorganisms (which the FDA threshold has accounted for). Based onthese results both of these antiseptic formulations, even as compared toa currently available product, produce significant (statistically andclinically) reductions in microorganisms load, even after repeatedexposure, as would be expected in a certain settings, such as forexample, hospitals, nursing homes, etc.

Example 7 Bacteria-Eliminating Efficacy of Sanitizing Formulations onAdult Hands

The present example was conducted to determine the antimicrobialeffectiveness of two test products (EtOH Antiseptic and AntisepticLotion) using a modification of the standardized test method ASTME2755-10, Determining the Bacteria Eliminating Effectiveness of HandSanitizer Formulations Using Hands of Adults.

A total of six subjects were tested, 3 per test material, over thecourse of 2 consecutive hand contaminations, the first followed by asample for baseline, and the second by a product application. As withthe Example 6, the indicator microorganism was S. marcescens (ATCC#14756). Mean log₁₀ reductions of S. marcescens was the basis forassessing the antimicrobial effectiveness of each test product.

A stock culture of Serratia marcescens (ATCC #14756) was prepared byaseptically transferring a lyophilized pellet to approximately 5.0 mL ofsterile TSB, which will then be incubated at 35° C.±2° C. for 25 hours±1hour. Two 500-mL flasks, each containing approximately 125 mL of TSB,were inoculated with 1.0 mL of the 24-hour broth culture, placed on aplatform shaker set at approximately 250 rpm, and incubated for 25hours±1 hour at 35° C.±2° C. 10.0-mL aliquots from each flask weredispensed into sterile graduated centrifuge tubes and centrifuged at4750 rpm±50 rpm for 30 minutes or until sedimentation was complete. Thesupernatant was decanted, and the pellet brought back to a volume of 1.0mL using TSB. The tubes were vortexed thoroughly and pooled into onecontainer prior to use. Prior to any withdrawal of culture, whether forhand contamination or for numbers assay, the suspension was stirredand/or swirled. The suspension was assayed for number of organisms atthe beginning and end of the use period. A suspension was not used formore than 8 hours.

Subjects were assigned randomly to use of one of the two test products(e.g., each person was assigned to use one, and only one, of the twotest products). Subjects' fingernails clipped were to a free edge of ˜1mm. All jewelry was removed from the hands and arms prior to theinception of the study period. Using tap water, a practicehand-inoculation procedure was performed by each subject, to reducesubject-to-subject variation do to application procedures. A 0.2-mLaliquot of tap water was transferred into a subject's cupped hands. Thetap water was then distributed evenly over both hands, not reachingabove the wrists, via gentle continuous massage.

A 30-second handwash using nonmedicated soap and a 30-second rinse wasperformed to remove dirt and oil from the hands prior to inception ofthe inoculation experiment. The temperature of the water used for thisand subsequent wash procedures was controlled at 40° C.±2° C.

Thereafter, a 0.2-mL aliquot of the inoculum suspension containingapproximately 1×10¹⁰ CFU/mL S. marcescens was transferred into thesubject's cupped hands. The inoculum was distributed evenly over bothhands, not reaching above the wrists, via continuous massage for 30seconds±5 seconds. After the timed 30-second massage, the Glove JuiceSampling Procedure was performed.

Within 5 minutes after contamination for baseline and after the singleproduct application, powder-free, sterile latex gloves were placed onsubjects' hands, and 75.0 mL Stripping Suspending Fluid (SSF) wasinstilled into each of the gloves. The wrists were secured, andattendants massaged the hands through the gloves in a standardizedmanner for 60 seconds. Following sampling a 5.0-mL aliquot of the glovejuice was removed from each of the gloves and separately diluted in 5.0mL Butterfield's Phosphate Buffer Solution with product neutralizers(BBP++) (dilution 10⁰). The 10⁰ dilutions then were serially diluted inBBP++, as appropriate. This first contamination cycle provided thebaseline population level. It was followed by a 30-second handwash usingnonmedicated soap and a 30-second rinse. Upon completion of thenonmedicated soap wash, subjects were required to wait a minimum of 5minutes prior to the start of the next contamination/product applicationcycle.

For the subsequent contamination, a 0.2-mL aliquot of the inoculumsuspension was again evenly distributed over both hands, as discussedabove. Thereafter, 0.4 mL of either the EtOH Antiseptic or theAntiseptic Lotion was dispensed into subjects' cupped, dry hands(notably, this is a significantly smaller application volume as comparedto other commercial formulations). The subjects rubbed the test productover the entire surface of the hands and fingers paying specialattention to nails and nail beds, and continued to rub into the skinuntil dry (as discussed above). Prior to any sample, subjects waited anadditional 5 minutes after the relevant product appeared dry on theskin. Thereafter, samples to evaluate the microbial reduction wereobtained using the Glove Juice Sampling Procedure (discussed above).

Samples were plated on duplicate spiral plates prepared from appropriatedilutions using Tryptic Soy Agar with product neutralizers (TSA+). Theplates were incubated at 25° C.±2° C. for approximately 48 hours, oruntil sufficient growth was observed. S. marcescens produces redcolonies, and only those colonies were counted. Colonies were countedand data recorded using the computerized Q-COUNT plate-counting system.If 10⁰ spiral plates gave an average count of zero, the average platecount was expressed as 1.00×10¹. A neutralization study (data not shown)assured that the neutralizers used in the recovery medium effectivelyquenched the antimicrobial activity of the test materials and were nottoxic to the challenge species. Study procedures were based on amodification of ASTM E2755-10, Determining the Bacteria-EliminatingEffectiveness of Hand Sanitizer Formulations Using Hands of Adults.R-values (estimated Log₁₀ number of viable microorganisms recovered fromeach subject's hand) was calculated as discussed in Example 6).

Log₁₀ reductions from baseline populations recovered from each of asubject's hands were calculated by subtracting the log₁₀ number ofviable microorganisms recovered from the hands following productapplication from the log₁₀ number of viable microorganisms recovered forbaseline. Statistical calculations of mean and standard deviation weregenerated on the log₁₀ data from baseline samples, post-productapplication samples, and the reductions from baseline. The statisticalresults are presented in Tables 17-22.

TABLE 17 Statistical Data of the Mean Log₁₀ Microbial Recoveries of S.marcescens and Mean Log₁₀ Microbial Reductions from Baseline FollowingSubjects' Use of 0.4 mL of EtOH Antiseptic Sample Minimum Maximum SampleSize Mean Value Value Baseline Log₁₀ Microbial 6 8.45 7.55 8.98 RecoveryPost-product Log₁₀ 6 6.30 5.16 7.72 Microbial Recovery Post-ProductApplication Log₁₀ 6 2.15 1.26 2.57 Reduction from Baseline

TABLE 18 Log₁₀ Microbial Recoveries of S. marcescens By Subject and HandFollowing Subjects' Use of 0.4 mL of EtOH Antiseptic Post-ProductBaseline Log₁₀ Post-product Log₁₀ Application Log₁₀ Microbial MicrobialReduction from Subject Hand Recovery Recovery Baseline 8 Left 8.98 7.721.26 Right 8.79 6.49 2.30 5 Left 7.82 5.25 2.57 Right 7.55 5.16 2.39 12Left 8.78 6.45 2.33 Right 8.80 6.75 2.05

TABLE 19 Statistical Data of the Mean Log₁₀ Microbial Recoveries of S.marcescens and Mean Log₁₀ Microbial Reductions from Baseline FollowingSubjects' Use of 0.4 mL of Antiseptic Lotion Sample Minimum MaximumSample Size Mean Value Value Baseline Log₁₀ Microbial 6 8.33 7.90 8.64Recovery Post-product Log₁₀ 6 6.86 6.39 7.61 Microbial RecoveryPost-Product Application Log₁₀ 6 1.47 1.03 1.83 Reduction from Baseline

TABLE 20 Log₁₀ Microbial Recoveries of S. marcescens By Subject and HandFollowing Subjects' Use of 0.4 mL of Antiseptic Lotion Post-ProductBaseline Log₁₀ Post-product Log₁₀ Application Log₁₀ Microbial MicrobialReduction from Subject Hand Recovery Recovery Baseline 8 Left 7.90 6.391.51 Right 8.42 6.60 1.83 5 Left 8.58 7.29 1.29 Right 8.64 7.61 1.03 12Left 8.29 6.62 1.67 Right 8.14 6.64 1.50

To ensure that the reductions detected were due to product applicationand not to falling microbial population numbers, the populations of theS. marcescens inoculum suspensions used in testing were over the testingperiod and are presented in Table 21.

TABLE 21 Initial and Final Populations of the S. marcescens InoculumSuspension Initial Population Final Population Test Date Group (CFU/mL)(CFU/mL) Apr. 9, 2013 1 (PM) 9.31 × 10⁹ 1.09 × 10⁹

Summary statistics were also generated on the log₁₀ reductions frombaseline for the EtOH Antiseptic, the Antiseptic Lotion and arepresented in Table 22.

TABLE 22 Statistical Summary Log₁₀ Microbial Reductions from Baseline95% Std. Dev. N Mean Min Max Std. Dev. Conf. Int. EtOH Antiseptic 62.150 1.262 2.568 0.466 0.273134-1.33272 Antiseptic Lotion 6 1.471 1.0281.826 0.282 0.165332-0.80672

A one-way ANOVA was performed to compare the efficacy of the EtOHAntiseptic and the Antiseptic Lotion. The results of the F-test(calculations not shown) indicated that the two antiseptics werestatistically distinct (p value of 0.012, with p<0.05 beingsignificant). With the sample size of 6, the 95% Confidence Intervals ofthe Standard Deviation were calculated (see Table 22) and are depictedgraphically in FIG. 4. Based on the overlapping 95% confidenceintervals, both products were the same in this evaluation, thereforepost-hoc analysis (the Tukey Method) was applied. Based on the post-hocanalysis, the EtOH antiseptic was found to result in greaterantimicrobial effects. However, depending on the microorganism tested,in several embodiments the two formulations may perform equivalently,and in still additional embodiments, the Antiseptic Lotion mayoutperform the EtOH Antiseptic. In summary, however, these datademonstrate that both the alcohol-based antiseptic and the water basedantiseptic lotion, according to several formulations disclosed herein,result in substantial reductions in bacterial load with just a singleapplication. These results indicate that, according to severalembodiments, even a single application of can yield significantantimicrobial effects. This efficacy is particularly important in manyenvironments in which, for example, repeated application is inefficientor impossible. In such cases, the efficacy of the disclosed formulationsensure a significant anti-microbial effect, even with a singleapplication, which may be the difference between stoppage andtransmission of microorganisms. Moreover, these results are particularlyunexpected in view of the reduced volume (0.4 mL) of the formulationsthat were used. Thus, in several embodiments, the efficacy of theformulations disclosed herein is sufficient to allow smaller volumes ofuse. In several embodiments, this results in better user compliance. Inseveral embodiments, this also reduces costs, without sacrificingefficacy.

Example 8 Extended Antimicrobial Efficacy of Sanitizing Formulations

As discussed above, several embodiments of the formulations disclosedherein result in rapid and broad-efficacy antimicrobial effects.However, many currently available and standard alcohol sanitizers andantibacterial soaps do not provide extended protection against microbes.For example, once the alcohol evaporates or the soap is rinsed off,hands can immediate become re-contaminated when they make contact with acontaminated surface. This presents a significant limitation toeffective hand hygiene, as even in healthcare settings, highly trainedworkers only comply with hand hygiene protocol 40% of the time withalcohol sanitizers, and compliance with handwashing is even lower. Thus,several embodiments of the formulations disclosed herein are designedwith persistence as well as efficacy in mind, in order to maximize theantimicrobial effect achieved when workers (or other types of users)utilize a sanitizing formulation, even if that use is less frequent thanrecommendations or in some embodiments, just a single application. Thepresent example tested products for both immediate kill and continuedprotection of the hands against transient bacteria. The products testedwere alcohol-based Antiseptic (comprising, according to one embodiment,70% ethanol w/v, EtOH Antiseptic), an Antiseptic Lotion (comprising,according to one embodiment, 0.2% BZT; Antiseptic Lotion), and a FoamingHand Soap (comprising, according to one embodiment, 0.2% BZT, FoamingSoap).

The testing utilized a modification of the ASTM1882 method, whichemploys a standard pigskin model as a substitute for human skin. Theproducts were applied as they would be used, according to severalembodiments, with the EtOH Antiseptic and Antiseptic Lotion tested asleave-on products and the Foaming Hand Soap lathered between two pigskinpieces and then rinsed clean. Testing was performed againstStaphylococcus aureus and Escherichia coli to test against both agram-positive and gram-negative bacteria, respectively.

The modified ASTM method 1882 was used, as detailed below. All controland test samples were performed in triplicate. Pigskin samples werecleaned with 70% ethanol and allowed to dry. A control was performed byrubbing a mild non-antibacterial soap (e.g., Ivory Soap) on a 4×4 cm²portion of pigskin for 20 seconds, then rinsing clean the pigskin andpatting it dry. The EtOH Antiseptic and Antiseptic Lotion were rubbed onindividual 4×4 cm² portions of pigskin for 20 seconds and allowed to dryon the pigskin. The Foaming Soap was applied, rubbed between two pigskinsamples, and then rinsed completely off and patted dry. After being leftundisturbed for a pre-determined sampling time (2 minutes, 1 hour, 4hours, and 8 hours (8 hours not tested for Foaming Soap in thisExample)), the treated pigskin was then pressed onto an inoculated platecontaining either S. aureus or E. coli organisms for 5 seconds, thenpressed onto a fresh, un-inoculated test plate for 5 seconds. Alltesting was performed in triplicate. The un-inoculated test plates wereincubated overnight (e.g., about 10-20 hours) at about 37° C. andcolonies were counted the following day.

The results for the antimicrobial effects of the EtOH Antiseptic againstS. aureus or E. coli are shown in Table 23 and FIG. 5.

TABLE 23 Persistent Antimicrobial Efficacy of EtOH Antiseptic S. AureusE. Coli Percent S. Aureus log Percent E. Coli log Time Point Reduction*Reduction Reduction** Reduction 2 minutes 99 1.87 98 1.65 1 hour 99 2.1497 1.34 4 hours 97 1.62 93 1.15 8 hours 90 0.99 89 0.95 *Initialinoculum of S. aureus of 2 × 10³ **Initial inoculum of E. coli of 5 ×10³

The results for the antimicrobial effects of the Antiseptic Lotionagainst S. aureus or E. coli are shown in Table 24 and FIG. 6.

TABLE 24 Persistent Antimicrobial Efficacy of Antiseptic Lotion S.Aureus E. Coli Percent S. Aureus log Percent E. Coli log Time PointReduction* Reduction Reduction** Reduction 2 minutes 97 1.6 98 1.80 1hour 98 1.85 98 1.71 4 hours 93 1.16 88 0.92 8 hours 85 0.84 89 0.98*Initial inoculum of S. aureus of 2 × 10³ **Initial inoculum of E. coliof 5 × 10³

The results for the antimicrobial effects of the Foaming Soap against S.aureus are shown in Table 25.

TABLE 25 Persistent Antimicrobial Efficacy of Foaming Soap S. Aureus S.Aureus Time Point Percent Reduction* log Reduction 2 minutes 97.6 1.63 1hour 97.8 1.67 4 hours 95.4 1.34 *Initial inoculum of S. aureus of 2 ×10³

These data demonstrate that each of the formulations tested forpersistence against S. aureus and/or E. coli exhibited substantialextended antimicrobial activity. Little loss of efficacy was detected inany of the products, even for up to eight hours after application, whichlikely extends well beyond the period in time in which a subject's handor hands would go without some method of cleansing or disinfecting.There was no notable loss in efficacy between the rapid-kill time framestested (see e.g., Example 5 above) and the 2 minute time point tested,and the reduction of S. aureus or E. Coli at 2 minutes was significant.Moreover, the EtOH Antiseptic and Antiseptic Lotion had only nominalreductions in efficacy after 1 hour, and only a 2-10% drop-off after 4hours. The Foaming Hand Soap, even after it had been rinsed from thepigskin model, showed similar efficacy, with no drop in performance anhour after application, and only a 2% drop after four hours. Carried outto 8 hours, the EtOH Antiseptic and the Antiseptic Lotion showed aslightly reduced efficacy of 90% reduction and 85% reduction in S.aureus (respectively) and an 89% reduction in E. coli. The long termefficacy of each of these formulations (e.g., 4 hours and longer) isparticularly unexpected given that the products are not directlycontacted with the bacteria (at least in the same fashion as in therapid-kill test), but rather the surface of the skin is “pre-treated”.Thus, in several embodiments, the application of one or more of theformulations disclosed herein functions in a prophylactic fashion, notonly killing microorganisms present on the surface (e.g., the hand) butalso reducing the likelihood of microorganisms re-colonizing thatsurface. Such preventative effects aid in reducing transmission ofmicroorganisms and thereby decrease the chance of spreading infectionfrom person to person (either directly or via contact with anothersurface).

The present example used a standardized method, thereby allowingretrospective comparison with the persistence of several alternativecommercial products. In 2006, a study demonstrated that (again with thesame protocol) Avagard (3M), Prevacare (J&J) and Triseptin (Healthpoint)showed efficacy ranging from 17%-67% reductions in transient bacteriawhen tested at 20 or 35 minutes. Surprisingly, the formulations testedin the present example all demonstrate more than 97% reduction inmicroorganisms at 1 hour and 88% (or more) at 4 hours (Shintre et al.,Int. J. of Hygiene and Environ. Health, 2006).

Thus, the formulations according to several embodiments herein exhibitsignificant antimicrobial properties against both gram-positive andgram-negative, transient bacteria at time points up to eight hours afterinitial application. This extended efficacy unexpected given that thisis based on a single application (and in the case of the Foaming Soap, asingle application followed by rinsing). This extended protection, inseveral embodiments, may be clinically significant, as circumstances maylimit the adherence to hand hygiene protocols. However the formulationsdisclosed herein, based at least in part on their persistent effects,may significantly reduce the transmission of pathogens and illness,thereby lowering infection rates in, for example, schools, shoppingareas or healthcare facilities, as well as in the general population andvarious other environments where microorganism colonization and/ortransmission could occur.

Example 9 Comparison of Extended Antimicrobial Efficacy Foaming SoapVersus Standard Soaps

Building on the data above related to the extended efficacy of severalof the formulations disclosed herein, the present example was performedto compare the long-term antimicrobial activity of the Foaming Soapaccording to several embodiments disclosed herein against other commonsoaps on the market. The soaps tested were Foaming Soap as disclosedherein, comprising benzethonium Chloride, 0.2% BZT, Brand 2antibacterial soap comprising 0.2% benzalkonium chloride, anantibacterial scrub comprising 0.4% chlorhexidine gluconate, andLifebuoy, a soap comprising phenol as an antibacterial agent.

Testing was performed at 2 minutes, 1 hour, and 6 hours afterapplication of the various soaps to determine how the persistent effectof the products changed with time. As discussed in Example 8 above, thetesting utilized a modification of the ASTM 1882 method, using pigskinas a substitute for human skin. The products were applied as they wouldbe used, with the soaps rinsed off after application. Testing wasperformed against Escherichia coli.

Pigskin samples were cleaned with 70% ethanol and allowed to dry. Acontrol was performed by rubbing a mild non-antibacterial soap (e.g.,Ivory Soap) on a 4×4 cm² portion of pigskin for 20 seconds, then rinsingclean the pigskin and patting it dry. The various soaps were applied totwo pigskin samples, which were rubbed together for 20 seconds, and thenrinsed completely off and patted dry. After being left undisturbed for apre-determined sampling time (2 minutes, 1 hour, 6 hours), the treatedpigskin was then pressed onto an inoculated plate containing E. coliorganisms for 5 seconds, then pressed onto a fresh, un-inoculated testplate for 5 seconds. All testing was performed in triplicate. Theun-inoculated test plates were incubated overnight (e.g., about 10-20hours) at about 37° C. and colonies were counted the following day.

The results for the comparison of persistent antimicrobial effects ofthe various soaps is shown in Table 26 and FIG. 7.

TABLE 26 Persistent Antimicrobial Efficacy of Soaps Against E. Coli.Foaming Soap 0.1% BZK Soap 4% CHG Scrub Phenol Time Percent log Percentlog Percent log Percent log Point Reduction Reduction ReductionReduction Reduction Reduction Reduction Reduction 2 minutes 100 3.55 5.00.03 100 3.6 44 0.27 1 hour 100 3.63 4.5 0.02 70 0.53 36 0.20 6 hours 820.74 5.0 0.04 70 0.53 33 0.17 * Initial inoculum of E. coli of 5 × 10³

These results demonstrate that, while several soaps offer modest to goodantimicrobial efficacy in the short term, the Foaming Soap formulation,as disclosed herein, significantly outperforms the other soaps tested atboth 1 hour and 6 hours post-application. Thus, as discussed above, theFoaming Soap not only offers the benefits of a short-term (e.g., rapid)antimicrobial effect, but also the longer term persistent effect, which(unexpectedly in view of the product being washed off) aids inpreventing of re-growth of any remaining microorganisms as well aslimiting the re-colonization of the surface with additionalmicroorganisms.

Example 10 Comparison of Efficacy of Soap Formulations AgainstDrug-Resistant Bacteria

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible forseveral difficult-to-treat infections in humans. MRSA are strains of S.aureus that have developed, resistance to beta-lactam antibiotics, suchas penicillins and cephalosporins. While the resistance itself does notequate with higher degree of virulence as compared to sensitive strains,the resistance does make MRSA infection more difficult to treat withstandard types of antibiotic. MRSA infections are present particularissues in hospitals, nursing homes, and other similar environments whereperson to person contact is high and individuals may have one or more ofopen wounds, invasive devices, and/or weakened immune systems. Incombination, these can present a population of individuals that are atgreater risk of infection than the general public, and therefore properhygiene to reduce and/or prevent infections, in particular MRSAinfections, is of paramount importance.

The present study, therefore, was performed to assess the efficacy ofcertain sanitizing formulations as disclosed herein against MRSA. Theprotocol was as performed in Example 9 above, in brief, pigskin sampleswere cleaned with 70% ethanol and allowed to dry. A control wasperformed by rubbing a mild non-antibacterial soap (e.g., Ivory Soap) ona 4×4 cm² portion of pigskin for 20 seconds, then rinsing clean thepigskin and patting it dry. The various soaps were applied to twopigskin samples, which were rubbed together for 20 seconds, and thenrinsed completely off and patted dry. After being left undisturbed for apre-determined sampling time (2 minutes, 1 hour, and 6 hours), thetreated pigskin was then pressed onto an inoculated plate containingMRSA for 5 seconds, then pressed onto a fresh, un-inoculated test platefor 5 seconds. All testing was performed in triplicate. Theun-inoculated test plates were incubated overnight (e.g., about 10-20hours) at about 37° C. and colonies were counted the following day.

The soaps tested were Foaming Soap as disclosed herein, comprisingbenzethonium Chloride, 0.2% BZT, Brand 1 antibacterial soap comprisingtriclosan; Brand 2 antibacterial soap comprising 0.2% benzalkoniumchloride, an antibacterial scrub comprising 0.4% chlorhexidinegluconate, and Lifebuoy, a soap comprising phenol as an antibacterialagent. The results from these experiments are summarized in Table 27 andshown in FIG. 8.

TABLE 27 Antimicrobial Efficacy of Soaps Against MRSA Product 2 minutes1 hour 6 hours Foaming Soap % Reduction 99.8 93.4 83.5 Log Reduction2.68 1.26 0.78 Brand 1 Soap with % Reduction 100 100 100 Triclosan LogReduction 3.70 3.76 3.78 0.1% BZK Soap % Reduction 64.7 56.7 32.9 LogReduction 0.5 0.46 0.18 4% CHG Scrub % Reduction 99.7 79.7 76.1 LogReduction 2.54 0.69 0.62 Phenol Soap % Reduction 58.8 50.9 40.6 LogReduction 0.4 0.31 0.23

These data demonstrate that the Foaming Soap, according to severalembodiments disclosed herein, significantly outperforms severalantibacterial products presently available, in particular whenconsidering the duration of antimicrobial effects. For example, theFoaming Soap killed nearly 100% of the MRSA at 2 minutes, with theefficacy only slightly diminishing at 1 hour. At 6 hours, the FoamingSoap still was able to reduce the MRSA by over 80%. In contrast, soapwith 0.1% BZK and phenol-containing soap achieved only 64.7% and 58.8%reduction in MRSA at 2 minutes. This efficacy is not only substantiallyless than the Foaming Soap, but it also is reduced over time, with the0.1% BZK formulation achieving only a 32.9% reduction at 6 hours, andthe phenol-containing soap achieving only 40.6 reduction in MRSA at 6hrs. The scrub containing 4% chlorhexidine gluconate results insubstantially similar antimicrobial effects at 2 minutes (99.7 versus99.8 for the Foaming Soap), however, like the above-mentioned products,the efficacy of the 4% CHG scrub drops off significantly by 1 hour (to79.7% reduction). At six hours post-application, the 4% CHG scrubachieved a 76.1% reduction, which is improved as compared to theabove-mentioned products, but is still less than the reduction achievedby the Foaming Soap. The Foaming Soap was not as effective as the Brand1 soap containing triclosan, which had 100% reduction at all timestested. However, as discussed above, the foaming soap formulations are,in several embodiments, triclosan-free, which, based on the data above,advantageously allows efficacious antimicrobial effects without theadverse effects associated with triclosan. Moreover, the Foaming Soapprovides an unexpectedly robust effect against MRSA, particularly for anextended duration, and with a reduced, and in several embodiments,nonexistent, risk of the development of microbial resistance to theefficacy of the formulation.

Example 11 Comparison of Efficacy of Leave-On Antiseptic FormulationsAgainst Drug-Resistant Bacteria

As discussed above, MRSA infections present a particular problem incertain environments, such as hospitals, nursing homes, etc., because ofthe more difficult regime needed to combat the infections. Preventingthe development and/or spread of infections would reduce the associatedmorbidity and mortality related to MRSA infections.

The present study, therefore, was performed to assess the efficacy ofcertain leave on sanitizing formulations as disclosed herein againstMRSA (as opposed to the soaps tested in Example 10, which are used andthen removed by rinsing). The protocol was as performed in Example 9above. The EtOH Antiseptic and Antiseptic Lotion were rubbed onindividual 4×4 cm² portions of pigskin for 20 seconds and allowed to dryon the pigskin. After being left undisturbed for a pre-determinedsampling time (2 minutes, 1 hour, or 6 hours the treated pigskin wasthen pressed onto an inoculated plate containing MRSA for 5 seconds,then pressed onto a fresh, un-inoculated test plate for 5 seconds. Alltesting was performed in triplicate. The un-inoculated test plates wereincubated overnight (e.g., about 10-20 hours) at about 37° C. andcolonies were counted the following day. The results from theseexperiments are summarized in Table 28 and depicted in FIG. 9.

TABLE 28 Antimicrobial Efficacy of Leave-on Sanitizing FormulationsAgainst MRSA Product 2 minutes 1 hour 6 hours EtOH Antiseptic %Reduction 99.4 94.4 76.3 Log Reduction 1.85 1.26 0.63 Antiseptic Lotion% Reduction 96.4 92.0 90 Log Reduction 1.45 1.08 1.02

These data demonstrate that the alcohol-based antiseptic and theantiseptic lotion, according to several embodiments disclosed herein,achieve considerable antimicrobial effects against MRSA, even atextended elapsed times after application. Both formulations were nearly100% effective at reducing MRSA at 2 minutes, with the efficacy onlyslightly diminishing at 1 hour. At 6 hours, the alcohol-based antisepticstill achieves a 76.3 percent reduction in MRSA. This may be a result ofthe evaporation of the alcohol during the elapsed 6 hours betweenapplication of the formulation and contact with the MRSA. However, giventhe significant morbidity and mortality related to MRSA infection andtransmission, this reduction at 6 hours may still have clinicalrelevance. The Antiseptic Lotion was still able to reduce MRSA by 90% at6 hours, which, in several embodiments, advantageously allows theAntiseptic Lotion to reduce risk of MRSA infection/transmission over anextended period of time. This persistence (of both formulations) mayhelp compensate for situations in which there is a risk of MRSAexposure, but recommended hygiene practices are not followed. Also, inseveral embodiments, these formulations achieve these effects withlimited risk of development of resistance by the MRSA.

Example 12 Comparison of Efficacy of Antiseptic Formulations AgainstNorovirus

As discussed above, various bacterial or viral outbreaks in certainenvironments can cause widespread illness. For example, human norovirusis one of the most common causes of acute viral gastroenteritis, causingabout 6 million clinical cases worldwide, including nearly 200,000deaths annually. Cruise ships, schools, health care settings, and othersettings in which individuals are in close and or repeated contact withone another present environments where transmission and spread ofinfection can occur rapidly.

The study described below will evaluate the virucidal efficacy ofseveral of the sanitizing formulations disclosed herein as compared to acontrol when the formulations are challenged with a human Norovirussurrogate. One such surrogate is Feline Calicivirus strain F9 (FCV; ATCC#VR-782). Human subjects are to be used in this study. The methodologyfor this is to be based on the Standard Test Method for Determining theVirus-Eliminating Effectiveness of Hygienic Handwash and Handrub AgentsUsing the Fingerpads of Adults, ASTM E 1838-10.

Feline kidney cells, such as for example ATCC #CCL-94 (or other suitablehost cells) are to be maintained as mono layers. Cells will be grown toapproximately 90% confluency and are to be less than 48 hours old beforebeing inoculation with virus. On the day of use, aliquots of the stockFCV virus are to be prepared with 5% Fetal Bovine Serum (FBS) as anOrganic Soil Load. Hard water (to be prepared according to acceptedStandard Operating Procedures) is to be used as a control. Controlevaluations of virus susceptibility (test sensitivity of cells to virus)will be performed. Control evaluations of the test/product neutralizer(e.g., neutralizer effect on cell susceptibility to virus) will beperformed.

Each subject will be in testing for 3 to 4 hours on a single day.Subjects will clip their fingernails to a free edge of approximately 1mm and all jewelry is to be removed from the hands and arms prior towashing. Subjects will use protective garments and a face shield. Eachsubject will perform a 30-second handwash using a nonmedicated soap toremove dirt and oil from the hands, which will be followed by a30-second rinse. During this handwash, the subjects will not be allowedto touch any part of the sink. The subjects will then dry their handsusing a clean paper towel and will receive into their cupped hands 5milliliters of 70% ethanol. The subjects will distribute the over thesurfaces of both of their hands by rubbing them together until dry.

Input Control

The thumbpads of each subject are to be demarcated, by, for example,pressing them to the open ends of screw-cap vials. This procedure willleave circular imprints on the thumbpads outlining the target areas forexposure. Each thumbpad will be contaminated with virus suspension(e.g., ˜10 μL) and will be eluted immediately using aneutralizer/elution solution. Maintenance Medium with 0.1% Tween 80 willbe used as the neutralizer/elution solution. The screw-cap vialscontaining 1.0 mL of the neutralizer/elution solution will be invertedover the contaminated thumbpads and will be held for approximately 5seconds. The screw-cap vials will then be repeatedly inverted and heldfor 5 seconds 20 times in succession. The mouths of the screw-cap vialswill be removed from the thumbpads, scraping the inside lips of thevials against the skin in an upward motion to recover as much fluid aspossible. The thumbpads will then be decontaminated by pressing them for3 minutes into a paper towel saturated with 5 mL of 70% ethanol,followed by an air-dry.

Baseline Control

Two randomly selected fingerpads (one from each hand) will be demarcatedas above. Each fingerpad will be contaminated with 10 μL of virussuspension and allowed to air-dry under ambient conditions. Eachfingerpad will then be placed over the mouth of a screw-cap vialcontaining 1.0 mL of the neutralizer/elution solution and sampled usingthe procedure described above (multiple, repeated inversions).

Test Material Efficacy

Two randomly selected fingerpads (one from each hand) will be demarcatedas above. Each designated fingerpad will be contaminated with 10 μL ofvirus suspension and allowed to air dry under ambient conditions. Eachvirus-contaminated fingerpad will be placed over the mouth of a vialcontaining test product and inverted. The test product will remain incontact with the contaminated area for approximately 10 seconds, and thevials will then be inverted 10 times in quick succession. Upon thecompletion of exposure, the mouths of the screw-cap vials will beremoved from the fingerpads. Each fingerpad will then be placed over themouth of a screw-cap vial containing 1.0 mL of the neutralizer/elutionsolution and will be sampled using the procedure described above.Controls (e.g., hard water) will be performed in the same fashion.

Results

In several embodiments, the antiseptic formulations tested will resultin substantial reduction in viral infectivity. In several embodiments,the reductions will be statistically greater than that achieved by thecontrol. In several embodiments, the reductions based on the applicationof the antiseptic formulations disclosed herein will be at least 2, willbe at least 3, will be at least 3.5, will be at least 4 (or greater)log₁₀ reductions in viral infectivity.

Although the embodiments of the inventions have been disclosed in thecontext of a certain preferred embodiments and examples, it will beunderstood by those skilled in the art that the present inventionsextend beyond the specifically disclosed embodiments to otheralternative embodiments and/or uses of the inventions and obviousmodifications and equivalents thereof. In addition, while a number ofvariations of the inventions have been shown and described in detail,other modifications, which are within the scope of the inventions, willbe readily apparent to those of skill in the art based upon thisdisclosure. It is also contemplated that various combinations orsubcombinations of the specific features and aspects of the embodimentsmay be made and still fall within one or more of the inventions.Further, the disclosure herein of any particular feature, aspect,method, property, characteristic, quality, attribute, element, or thelike in connection with an embodiment can be used in all otherembodiments set forth herein. Accordingly, it should be understood thatvarious features and aspects of the disclosed embodiments can becombined with or substituted for one another in order to form varyingmodes of the disclosed inventions. For all of the embodiments describedherein the steps of the methods need not be performed sequentially.Thus, it is intended that the scope of the inventions herein disclosedshould not be limited by the particular disclosed embodiments describedabove.

The ranges disclosed herein also encompass any and all overlap,sub-ranges, and combinations thereof. Language such as “up to,” “atleast,” “greater than,” “less than,” “between,” and the like includesthe number recited. Numbers preceded by a term such as “about” or“approximately” include the recited numbers. For example, “about 10micrograms” includes “10 micrograms.

1-20. (canceled)
 21. A foaming soap comprising, by weight: about 50.0%to about 65.0% water; about 5.0% to about 20.0% ethyl alcohol; about0.005% to about 0.1% polyquaternium-10; about 0.1% to about 2.0% citricacid; about 0.1% to about 2.0% zinc gluconate; about 0.05% to about 2.0%benzethonium chloride; and about 0.1% to about 1.0% farnesol.
 22. Thefoaming soap of claim 21, which comprises, by weight, about 10.0% ethylalcohol.
 23. The foaming soap of claim 21, which comprises, by weight,about 0.01% polyquaternium-10.
 24. The foaming soap of claim 21, whichcomprises, by weight, about 0.2% benzethonium chloride.
 25. The foamingsoap of claim 21, which comprises, by weight, about 0.5% farnesol. 26.The foaming soap of claim 21, which comprises, by weight, about 0.7%citric acid.
 27. The foaming soap of claim 21, which comprises, byweight, about 0.3% zinc gluconate.
 28. The foaming soap of claim 21,which comprises, by weight: about 50.0% to about 65.0% water; about10.0% ethyl alcohol; about 0.01% polyquaternium-10; about 0.7% citricacid; about 0.3% zinc gluconate; about 0.2% benzethonium chloride; andabout 0.5% farnesol.
 29. The foaming soap of claim 21, furthercomprising butylene glycol, lauramine oxide (LO), cocamiopropyl betaine,sodium chloride), cetrimonium chloride, PPG-2 hydroxyethyl cocamide,methylchloroisothiazolinone, glycerin, water aloe barbadensis leafextract, a fragrance, and polyaminopropyl biguanide.
 30. The foamingsoap of claim 29, which comprises, by weight, about 0.5% to about 2.0%butylene glycol.
 31. The foaming soap of claim 29, which comprises, byweight, about 5.0% to about 10.0% mackamine LO which comprises water andlauramine oxide.
 32. The foaming soap of claim 29, which comprises, byweight, about 5.0% to about 10.0% caltaine C-35 which comprises water,cocamiopropyl betaine, and sodium chloride.
 33. The foaming soap ofclaim 29, which comprises, by weight, about 3.0% to about 10.0%carsoquat CT-429 which comprises water and cetrimonium chloride.
 34. Thefoaming soap of claim 29, which comprises, by weight, about 1.0% toabout 5.0% PPG-2 hydroxyethyl cocamide.
 35. The foaming soap of claim29, which comprises, by weight, about 0.01% to about 1.0%methylchloroisothiazolinone.
 36. The foaming soap of claim 29, whichcomprises, by weight, about 0.05% to about 1.0% actiphyte of aloe verawhich comprises glycerin and water aloe barbadensis leaf extract. 37.The foaming soap of claim 29, which comprises, by weight, about 0.01% toabout 0.5% cosmocil CQ which comprises water and polyaminopropylbiguanide.
 38. The foaming soap of claim 21, which comprises, by weight:about 50.0% to about 65.0% water; about 5.0% to about 20.0% ethylalcohol; about 0.005% to about 0.1% polyquaternium-10; about 0.1% toabout 2.0% citric acid; about 0.1% to about 2.0% zinc gluconate; about0.05% to about 2.0% benzethonium chloride; about 0.1% to about 1.0%farnesol; about 0.005% to about 0.1% polyquaternium-10; about 0.1% toabout 2.0% citric acid; about 0.5% to about 2.0% butylene glycol; about0.1% to about 2.0% zinc gluconate; about 0.05% to about 2.0%benzethonium chloride; about 5.0% to about 10.0% mackamine LO; about5.0% to about 10.0% caltaine C-35; about 3.0% to about 10.0% carsoquatCT-429; about 1.0% to about 5.0% PPG-2 hydroxyethyl cocamide; about0.01% to about 1.0% methylchloroisothiazolinone; about 0.05% to about1.0% actiphyte of aloe vera; about 0.01 to about 0.10% of the fragrance;and about 0.01% to about 0.5% cosmocil CQ.